No description available.

Peroxisome proliferator-activated receptors (PPARs) are members of a large group of ligand-regulated transcription factors that includes nuclear receptors for steroid and thyroid hormones, retinoids and vitamin D���. Synthetic fibrates and thiazolidinediones that bind to and activate PPARs are used efficaciously in humans to remedy hypertriglyceridemia and non-insulin dependent diabetes mellitus, respectively. The objective of the studies described herein was to elucidate the molecular mechanisms of ligand-dependent PPAR signaling. Several PPAR ligands, including WY-14,643, were demonstrated to directly induce PPAR�� conformational changes as evidenced by a differential protease sensitivity assay. Conformational changes were induced in a dose-dependent manner which paralleled that of ligand to induce transcriptional activation. Direct interaction of ligands with, and the resulting conformational alterations in, PPAR�� may facilitate interaction of the receptor with transcriptional intermediary factors and thus may underlie the molecular basis of ligand-dependent transcriptional activation mediated by PPAR��. The yeast two hybrid screen was utilized to identify downstream components of the PPAR�� signaling pathway. Using this technique, the coactivator proteins, p300 and steroid receptor coactivator-1 (SRC-1), were identified as PPAR��-interacting proteins and WY-14,643 potentiated these interactions. p300 also enhanced the transcriptional activation properties of PPAR�� and, therefore, can be considered a bona fide coactivator for this nuclear receptor. Nuclear receptor corepressor (NCoR) was also isolated as a PPAR��-interacting protein from a yeast two hybrid screen. In contrast to the ligand enhanced PPAR��-coactivator interactions, WY-14,643 inhibited NCoR interaction with PPAR��. NCoR and the coactivators, p300 and SRC-1, were also demonstrated to require distinct receptor regions for efficient interaction with PPAR��. Results described herein demonstrate that ligand induces PPAR�� conformational changes, promotes PPAR��-coactivator (p300 and SRC-1) interactions, and inhibits PPAR��-NCoR interactions. We hypothesize that such molecular events are critical for ligand-dependent transcriptional activation by PPAR��. These results contribute additional knowledge as to the molecular mechanisms of PPAR-dependent signaling and may act as a starting part for the improvement and/or development of therapeutic strategies aimed at manipulating this signaling pathway. / Graduation date: 1999

M.S. / Molecular Biology / This thesis describes the cloning and analysis of PER6, a gene required for peroxisome biogenesis in Pichia pastoris. The gene was cloned by functional complementation of a per6 P. pastoris mutant strain that was one of a number of peroxisome-deficient mutants isolated in this laboratory. The complementing activity was localized to a small DNA fragment by subcloning and Northern filter hybridization analysis and the DNA sequence of the fragment was determined. The sequence revealed a 1296-bp open reading frame which potentially encodes a 432-amino acid protein of 49 kD. The gene was transcribed into a message of 1.4 kilobases that was constitutively expressed but induced several-fold in cells growing on methanol. A mutant strain with a deletion of a large portion of the open reading frame was constructed and used to genetically demonstrate that the cloned gene was identical to the defective gene in the originally isolated per6 mutant. The predicted amino acid sequence of the PER6 product revealed several interesting features, including a significant regional similarity to PAF-1, a gene known to be defective in some patients with Zellweger syndrome, a lethal human genetic disease caused by peroxisome deficiency. Finally, the PER6 product was produced in E. coli and purified to serve as antigen for antibody production.

Thesis (M.S.), Oregon Graduate Institute of Science & Technology, 1993.

Thesis (Ph.D.) - University of Queensland, 2004. / Includes bibliography.

Lo Kam Chun. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references (leaves 310-323). / Abstracts in English and Chinese. / Abstract --- p.i / Abstract (Chinese version) --- p.iii / Acknowledgements --- p.v / Table of Contents --- p.vi / List of Abbreviations --- p.xxvi / List of Unit Abbreviations --- p.xxviii / List of Figures --- p.xxix / List of Tables --- p.xxxviii / Chapter Chapter 1 --- Introduction --- p.1 / Chapter Chapter 2 --- Literature Review --- p.3 / Chapter 2.1 --- Peroxisome proliferator-activated receptors (PPARs) --- p.3 / Chapter 2.1.1 --- Structure of PPAR --- p.4 / Chapter 2.1.2 --- Ligands of PPAR --- p.6 / Chapter 2.1.3 --- Peroxisome proliferator responsive elements (PPREs) --- p.8 / Chapter 2.1.4 --- Coactivators and corepressors of PPAR --- p.8 / Chapter 2.1.5 --- Signaling cross-talk between PPAR and other nuclear hormone receptors --- p.11 / Chapter 2.1.6 --- Tissue distribution of PPAR --- p.12 / Chapter 2.2 --- PPARα and lipid metabolism --- p.13 / Chapter 2.3 --- "PPARα, cholesterol and lipoprotein metabolism" --- p.18 / Chapter 2.4 --- PPARα and glucose metabolism --- p.20 / Chapter 2.5 --- PPARα and amino acid metabolism --- p.21 / Chapter 2.6 --- "PPARα, atherosclerosis and inflammation" --- p.21 / Chapter 2.7 --- PPARα and epidermal differentiation --- p.23 / Chapter 2.8 --- PPARα and peroxisome proliferator-induced liver injury --- p.24 / Chapter Chapter 3 --- Objectives --- p.25 / Chapter Chapter 4 --- Materials and Methods --- p.26 / Chapter 4.1 --- Animal tail-genotyping --- p.26 / Chapter 4.1.1 --- Materials --- p.26 / Chapter 4.1.2 --- Methods --- p.26 / Chapter 4.2 --- Animal treatment --- p.29 / Chapter 4.2.1 --- Animals --- p.30 / Chapter 4.2.2 --- Treatment --- p.30 / Chapter 4.3 --- "Serum β-hydroxybutyrate, triglyceride and cholesterol analyses" --- p.30 / Chapter 4.3.1 --- Materials --- p.32 / Chapter 4.3.2 --- Methods --- p.32 / Chapter 4.3.2.1 --- Serum preparation --- p.32 / Chapter 4.3.2.2 --- β-hydroxybutyrate determination --- p.32 / Chapter 4.3.2.3 --- Triglyceride determination --- p.33 / Chapter 4.3.2.4 --- Cholesterol determination --- p.33 / Chapter 4.4 --- Total RNA isolation --- p.34 / Chapter 4.4.1 --- Materials --- p.34 / Chapter 4.4.2 --- Methods --- p.34 / Chapter 4.5 --- DNase I treatment --- p.35 / Chapter 4.5.1 --- Materials --- p.35 / Chapter 4.5.2 --- Methods --- p.35 / Chapter 4.6 --- Reverse transcription (RT) and non-fluorescent PCR (non-fluoroDD PCR) --- p.36 / Chapter 4.6.1 --- Materials --- p.41 / Chapter 4.6.2 --- Methods --- p.41 / Chapter 4.7 --- Reverse transcription (RT) and fluorescent PCR (fluoroDD PCR) --- p.42 / Chapter 4.7.1 --- Materials --- p.42 / Chapter 4.7.2 --- Methods --- p.42 / Chapter 4.8 --- Fluorescent differential display --- p.42 / Chapter 4.8.1 --- Materials --- p.43 / Chapter 4.8.2 --- Methods --- p.43 / Chapter 4.9 --- Excision of differentially expressed cDNA fragments --- p.44 / Chapter 4.9.1 --- Materials --- p.44 / Chapter 4.9.2 --- Methods --- p.44 / Chapter 4.10 --- Reamplification of excised cDNA fragments --- p.44 / Chapter 4.10.1 --- Materials --- p.45 / Chapter 4.10.2 --- Methods --- p.45 / Chapter 4.11 --- Subcloning of reamplified cDNA fragments --- p.46 / Chapter 4.11.1 --- Materials --- p.46 / Chapter 4.11.2 --- Methods --- p.46 / Chapter 4.12 --- Reverse dot blot analysis of subcloned cDNA fragments --- p.50 / Chapter 4.12.1 --- Materials --- p.50 / Chapter 4.12.2 --- Methods --- p.51 / Chapter 4.12.2.1 --- Mini-preparation of plasmid DNA from recombinant clones ´ؤ´ؤalkaline lysis method --- p.51 / Chapter 4.12.2.2 --- Dot blot preparation --- p.52 / Chapter 4.12.2.3 --- cDNA library and probe preparation --- p.52 / Chapter 4.12.2.4 --- Hybridization and signal detection --- p.54 / Chapter 4.13 --- Sequencing of reverse dot blot-confirmed cDNA fragments --- p.55 / Chapter 4.13.1 --- Materials --- p.55 / Chapter 4.13.2 --- Methods --- p.55 / Chapter 4.13.2.1 --- Mini-preparation of plasmid DNA from recombinant clones ´ؤ´ؤQiagen method --- p.55 / Chapter 4.13.2.2 --- DNA sequencing by CEQ dye terminator cycle sequencing --- p.56 / Chapter 4.13.2.3 --- BLAST search against computer database --- p.57 / Chapter 4.14 --- Northern blot analysis of sequenced cDNA fragments --- p.57 / Chapter 4.14.1 --- Materials --- p.58 / Chapter 4.14.2 --- Methods --- p.58 / Chapter 4.14.2.1 --- Midi-preparation of plasmid DNA from recombinant clones for probe preparation ´ؤ´ؤQiagen method --- p.58 / Chapter 4.14.2.2 --- DIG-labeled cDNA probe preparation --- p.59 / Chapter 4.14.2.3 --- Formaldehyde-agarose gel electrophoresis --- p.60 / Chapter 4.14.2.4 --- Hybridization and signal detection --- p.61 / Chapter Chapter 5 --- Results --- p.63 / Chapter 5.1 --- Confirmation of genotypes by PCR tail-genotyping --- p.63 / Chapter 5.2 --- Morphological observation --- p.63 / Chapter 5.3 --- "Serum β-hydroxybutyrate, triglyceride and cholesterol levels" --- p.67 / Chapter 5.4 --- RT and non-fluoroDD PCR --- p.69 / Chapter 5.5 --- RT and fluoroDD PCR --- p.94 / Chapter 5.6 --- Reamplification of cDNA fragments --- p.121 / Chapter 5.7 --- Subcloning of reamplified cDNA fragments --- p.131 / Chapter 5.8 --- Reverse dot blot analysis of subcloned cDNA fragments --- p.174 / Chapter 5.9 --- Sequencing of reverse dot blot-confirmed cDNA fragments --- p.243 / Chapter 5.10 --- Northern blot analysis of sequenced cDNA fragments --- p.262 / Chapter Chapter 6 --- Discussions --- p.282 / Chapter 6.1 --- PPARα and hepatic lipid metabolism during fasting --- p.282 / Chapter 6.2 --- Enlargement of liver in starved PPARα-null mice --- p.283 / Chapter 6.3 --- "Alteration of serum β-hydroxybutyrate, triglyceride and cholesterol levels by PPARa during fasting" --- p.284 / Chapter 6.4 --- Identification of PPARα-dependent and fasting- responsive genes by fluorescent differential display and PPARα-null mice --- p.285 / Chapter 6.4.1 --- Roles of PPARα-dependent and fasting-responsive genes --- p.289 / Chapter 6.4.1.1 --- Lipid metabolism --- p.290 / Chapter 6.4.1.2 --- Protein metabolism --- p.299 / Chapter 6.4.1.3 --- Other functions --- p.302 / Chapter Chapter 7 --- Conclusions --- p.307 / Chapter Chapter 8 --- Future Studies --- p.309 / Chapter 8.1 --- Further identification of fasting-responsive and PPARα- dependent genes --- p.309 / Chapter 8.2 --- Characterization of the mouse EST --- p.309 / References --- p.310 / Chapter Appendix A: --- Protocol for reaction preparation / Chapter Table A1 --- PCR-tail genotyping --- p.324 / Chapter Table A2 --- DNase I treatment of total RNA --- p.324 / Chapter Table A3 --- Reverse transcription (RT) for both non-fluoroDD and fluooDD PCR --- p.324 / Chapter Table A4 --- Non-fluoroDD PCR --- p.325 / Chapter Table A5 --- FluoroDD PCR --- p.325 / Chapter Table A6 --- Reamplification of cDNA fragments --- p.325 / Chapter Table A7 --- Reverse transcription in cDNA library synthesis for reverse dot blot analysis --- p.326 / Chapter Table A8 --- RT-PCR in cDNA library synthesis for reverse dot blot analysis --- p.326 / Chapter Appendix B: --- Phenol-chloroform extraction for recombinant subclones / Chapter Figure B1 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gel D --- p.327 / Chapter Figure B2 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gel E --- p.328 / Chapter Figure B3 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gels G and HH --- p.329 / Chapter Figure B4 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gel JJ --- p.330 / Chapter Figure B5 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gel LL --- p.331 / Chapter Figure B6 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gel MM --- p.332 / Chapter Figure B7 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gels NN and OO (fragments OO1 - OO4) --- p.333 / Chapter Figure B8 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gel 00 (fragments OO5 - OO7) --- p.334 / Chapter Figure B9 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gel OO (fragments OO8 and OO9) --- p.335 / Chapter Figure B10 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gel PP --- p.336 / Chapter Figure B11 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gel QQ (fragments QQ1 and QQ3) --- p.337 / Chapter Figure B12 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gel QQ (fragments QQ4 - QQ6) --- p.338 / Chapter Figure B13 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gel RR --- p.339 / Chapter Figure B14 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gels SS and TT --- p.340 / Chapter Figure B15 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gel UU --- p.341 / Chapter Figure B16 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gel VV --- p.342 / Chapter Figure B17 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gel WW --- p.343 / Chapter Figure B18 --- Phenol-chloroform extraction for recombinant subclones containing cDNA fragments excised from fluoroDD gel XX --- p.344 / Chapter Appendix C: --- DNA sequence and sequence alignment of cDNA subclones / Chapter C1.1 --- DNA sequence of cDNA subclone Dl#2 using Ml3 forward -20 primer and the sequence of 3´ة-AP3 --- p.345 / Chapter C1.2 --- Sequence alignment of cDNA subclone Dl#2 with mouse CYP4A10 by BLAST searching with the National Center for Biotechnology Information database --- p.345 / Chapter C1.3 --- Summary of sequence alignment of cDNA subclone Dl#2 with mouse CYP4A10 --- p.345 / Chapter C2.1 --- DNA sequence of cDNA subclone Dl#2 using Ml3 reverse primer and the sequence of 5´ة-ARP12 --- p.346 / Chapter C2.2 --- Sequence alignment of cDNA subclone Dl#2 with mouse CYP4A10 by BLAST searching with the National Center for Biotechnology Information database --- p.346 / Chapter C2.3 --- Summary of sequence alignment of cDNA subclone Dl#2 with mouse CYP4A10 --- p.346 / Chapter C3.1 --- DNA sequence of cDNA subclone D2#9 using Ml3 forward -20 primer and the sequence of 5'-ARP12 --- p.347 / Chapter C3.2 --- Sequence alignment of cDNA subclone D2#9 with mouse Bhmt by BLAST searching with the National Center for Biotechnology Information database --- p.347 / Chapter C3.3 --- Summary of sequence alignment of cDNA subclone D2#9 with mouse Bhmt --- p.347 / Chapter C4.1 --- DNA sequence of cDNA subclone D3#3 using Ml3 forward -20 primer and the sequence of 5'-ARP12 --- p.348 / Chapter C4.2 --- Sequence alignment of cDNA subclone D3#3 with mouse hypothetical protein MMT-7 by BLAST searching with the National Center for Biotechnology Information database --- p.348 / Chapter C4.3 --- Summary of sequence alignment of cDNA subclone D3#3 with mouse hypothetical protein MMT-7 --- p.348 / Chapter C5.1 --- DNA sequence of cDNA subclone D3#3 using M13 reverse primer and the sequence of 3´ة-AP3 --- p.349 / Chapter C5.2 --- Sequence alignment of cDNA subclone D3#3 with mouse hypothetical protein MMT-7 by BLAST searching with the National Center for Biotechnology Information database --- p.349 / Chapter C5.3 --- Summary of sequence alignment of cDNA subclone D3#3 with mouse hypothetical protein MMT-7 --- p.349 / Chapter C6.1 --- DNA sequence of cDNA subclone LL1#7 using Ml3 reverse primer and the sequence of 3´ة-AP4 --- p.350 / Chapter C6.2 --- Sequence alignment of cDNA subclone LL1#7 with mouse Ctsl by BLAST searching with the National Center for Biotechnology Information database --- p.350 / Chapter C6.3 --- Summary of sequence alignment of cDNA subclone LL1#7 with mouse Ctsl --- p.350 / Chapter C7.1 --- DNA sequence of cDNA subclone LL1#11 using Ml3 forward -20 primer and the sequence of 5'-ARP6 --- p.351 / Chapter C7.2 --- Sequence alignment of cDNA subclone LL1#11 with mouse Ctsl by BLAST searching with the National Center for Biotechnology Information database --- p.351 / Chapter C7.3 --- Summary of sequence alignment of cDNA subclone LL1#11 with mouse Ctsl --- p.351 / Chapter C8.1 --- DNA sequence of cDNA subclone MM1#1 using M13 forward -20primer and the sequence of 3'-AP2 --- p.352 / Chapter C8.2 --- Sequence alignment of cDNA subclone MM1#1 with mouse CYP4A10 by BLAST searching with the National Center for Biotechnology Information database --- p.352 / Chapter C8.3 --- Summary of sequence alignment of cDNA subclone MM1#1 with mouse CYP4A10 --- p.352 / Chapter C9.1 --- DNA sequence of cDNA subclone MM1#1 using Ml3 reverse primer and the sequence of 5´ة-ARP7 --- p.353 / Chapter C9.2 --- Sequence alignment of cDNA subclone MM1#1 with mouse CYP4A110 by BLAST searching with the National Center for Biotechnology Information database --- p.353 / Chapter C9.3 --- Summary of sequence alignment of cDNA subclone MM1#1 with mouse CYP4A10 --- p.353 / Chapter C10.1 --- DNA sequence of cDNA subclone 002#6 using Ml3 forward -20 primer and the sequence of 3'-AP9 --- p.354 / Chapter C10.2 --- Sequence alignment of cDNA subclone 002#6 with rat ASL by BLAST searching with the National Center for Biotechnology Information database --- p.354 / Chapter C10.3 --- Summary of sequence alignment of cDNA subclone 002#6 with rat ASL --- p.354 / Chapter C11.l --- DNA sequence of cDNA subclone 002#9 using Ml3 reverse primer and the sequence of 5'-ARP3 --- p.355 / Chapter C11.2 --- Sequence alignment of cDNA subclone 002#9 with rat ASL by BLAST searching with the National Center for Biotechnology Information database --- p.355 / Chapter C11.3 --- Summary of sequence alignment of cDNA subclone 002#9 with rat ASL --- p.355 / Chapter C12.1 --- DNA sequence of cDNA subclone 003#4 using Ml3 forward -20 primer and the sequence of 5'-ARP3 --- p.356 / Chapter C12.2 --- Sequence alignment of cDNA subclone O03#4 with mouse Phyh by BLAST searching with the National Center for Biotechnology Information database --- p.356 / Chapter C12.3 --- Summary of sequence alignment of cDNA subclone 003#4 with mouse Phyh --- p.356 / Chapter C13.1 --- DNA sequence of cDNA subclone 003#9 using Ml3 reverse primer and the sequence of 5'-ARP3 --- p.357 / Chapter C13.2 --- Sequence alignment of cDNA subclone 003#9 with rat ASL by BLAST searching with the National Center for Biotechnology Information database --- p.357 / Chapter C13.3 --- Summary of sequence alignment of cDNA subclone 003#9 with rat ASL --- p.357 / Chapter C14. --- DNA sequence of cDNA subclone 004#3 using M13 forward -20 primer and the sequence of 5'-ARP3 --- p.358 / Chapter C14.2 --- Sequence alignment of cDNA subclone 004#3 with rat ASL by BLAST searching with the National Center for Biotechnology Information database --- p.358 / Chapter C14.3 --- Summary of sequence alignment of cDNA subclone 004#3 with rat ASL --- p.358 / Chapter C15.1 --- DNA sequence of cDNA subclone 004#3 using Ml3 reverse primer and the sequence of 3'-AP9 --- p.359 / Chapter C15.2 --- Sequence alignment of cDNA subclone 004#3 with rat ASL by BLAST searching with the National Center for Biotechnology Information database --- p.359 / Chapter C15.3 --- Summary of sequence alignment of cDNA subclone 004#3 with rat ASL --- p.359 / Chapter C16.1 --- DNA sequence of cDNA subclone 004#4 using Ml 3 forward -20primer and the sequence of 5?-ARP3 --- p.360 / Chapter C16.2 --- Sequence alignment of cDNA subclone 004#4 with mouse CYP4A14 by BLAST searching with the National Center for Biotechnology Information database --- p.360 / Chapter C16.3 --- Summary of sequence alignment of cDNA subclone 004#4 with mouse CYP4A14 --- p.360 / Chapter C17.1 --- DNA sequence of cDNA subclone 004#4 using M13 reverse primer and the sequence of 3´ة-AP9 --- p.361 / Chapter C17.2 --- Sequence alignment of cDNA subclone 004#4 with mouse CYP4A14 by BLAST searching with the National Center for Biotechnology Information database --- p.361 / Chapter C17.3 --- Summary of sequence alignment of cDNA subclone 004#4 with mouse CYP4A14 --- p.361 / Chapter C18.1 --- DNA sequence of cDNA subclone 004#10 using M13 forward -20primer and the sequence of 3´ة-AP9 --- p.362 / Chapter C18.2 --- a Sequence alignment of cDNA subclone 004#10 with rat PBE by BLAST searching with the National Center for Biotechnology Information database --- p.362 / Chapter C18.3 --- a Summary of sequence alignment of cDNA subclone 004#10 with rat PBE --- p.362 / Chapter C18.2 --- b Sequence alignment of cDNA subclone 004#10 with mouse L-PBE by BLAST searching with the National Center for Biotechnology Information database --- p.363 / Chapter C18.3 --- b Summary of sequence alignment of cDNA subclone 004#10 with mouse L-PBE --- p.363 / Chapter C19.1 --- DNA sequence of cDNA subclone 005#13 using M13 forward -20primer and the sequence of 3'-AP9 --- p.364 / Chapter C19.2 --- Sequence alignment of cDNA subclone 005#13 with mouse CYP4A14 by BLAST searching with the National Center for Biotechnology Information database --- p.364 / Chapter C19.3 --- Summary of sequence alignment of cDNA subclone 005#13 with mouse CYP4A14 --- p.364 / Chapter C20.1 --- DNA sequence of cDNA subclone 005#13 using Ml3 reverse primer and the sequence of 5'-ARP3 --- p.365 / Chapter C20.2 --- Sequence alignment of cDNA subclone 005#13 with mouse CYP4A14 by BLAST searching with the National Center for Biotechnology Information database --- p.365 / Chapter C20.3 --- Summary of sequence alignment of cDNA subclone 005#13 with mouse CYP4A14 --- p.365 / Chapter C21.1 --- DNA sequence of cDNA subclone 006#2 using Ml3 forward -20 primer and the sequence of 3´ة-AP9 --- p.366 / Chapter C21.2 --- Sequence alignment of cDNA subclone 006#2 with mouse CYP4A14 by BLAST searching with the National Center for Biotechnology Information database --- p.366 / Chapter C21.3 --- Summary of sequence alignment of cDNA subclone 006#2 with mouse CYP4A14 --- p.366 / Chapter C22.1 --- DNA sequence of cDNA subclone 006#8 using Ml3 forward -20 primer and the sequence of 5'-ARP3 --- p.367 / Chapter C22.2 --- Sequence alignment of cDNA subclone 006#8 with mouse CYP4A14 by BLAST searching with the National Center for Biotechnology Information database --- p.367 / Chapter C22.3 --- Summary of sequence alignment of cDNA subclone 006#8 with mouse CYP4A14 --- p.367 / Chapter C23.1 --- DNA sequence of cDNA subclone 006#10 using Ml3 reverse primer and the sequence of 5'-ARP3 --- p.368 / Chapter C23.2 --- Sequence alignment of cDNA subclone 006#10 with mouse CYP4A14 by BLAST searching with the National --- p.368 / Chapter C23.3 --- Summary of sequence alignment of cDNA subclone 006#10 with mouse CYP4A14 --- p.368 / Chapter C24.1 --- DNA sequence of cDNA subclone 007#6 using Ml3 reverse primer and the sequence of 5'-ARP3 --- p.368 / Chapter C24.2 --- Sequence alignment of cDNA subclone 007#6 with mouse Spil-3 by BLAST searching with the National Center for Biotechnology Information database --- p.369 / Chapter C24.3 --- Summary of sequence alignment of cDNA subclone 007#6 with mouse Spil-3 --- p.369 / Chapter C25.1 --- DNA sequence of cDNA subclone 007#10 using Ml3 forward -20 primer and the sequence of 3'-AP9 --- p.370 / Chapter C25.2 --- Sequence alignment of cDNA subclone 007#10 with mouse Spil-3 by BLAST searching with the National Center for Biotechnology Information database --- p.370 / Chapter C25.3 --- Summary of sequence alignment of cDNA subclone 007#10 with mouse Spil-3 --- p.370 / Chapter C26.1 --- DNA sequence of cDNA subclone 008#10 using Ml3 forward -20 primer and the sequence of 5'-ARP3 --- p.371 / Chapter C26.2 --- Sequence alignment of cDNA subclone 008#10 with mouse Hpcl by BLAST searching with the National Center for Biotechnology Information database --- p.371 / Chapter C26.3 --- Summary of sequence alignment of cDNA subclone 008#l0 with mouse Hpcl --- p.371 / Chapter C27.1 --- DNA sequence of cDNA subclone 009#6 using Ml3 forward -20 primer and the 3´ة-AP9 --- p.372 / Chapter C27.2 --- Sequence alignment of cDNA subclone 009#6 with mouse Hpcl by BLAST searching with the National Center for Biotechnology Information database --- p.372 / Chapter C27.3 --- Summary of sequence alignment of cDNA subclone 009#6 with mouse Hpcl --- p.372 / Chapter C28.1 --- DNA sequence of cDNA subclone 009#8 using Ml3 reverse primer and the sequence of 3'-AP9 --- p.373 / Chapter C28.2 --- Sequence alignment of cDNA subclone 009#8 with mouse Hpcl by BLAST searching with the National Center for Biotechnology Information database --- p.373 / Chapter C28.3 --- Summary of sequence alignment of cDNA subclone 009#8 with mouse Hpcl --- p.373 / Chapter C29.1 --- DNA sequence of cDNA subclone PP3#3 using M13 forward -20 primer and the sequence of 5'-ARP3 --- p.374 / Chapter C29.2 --- Sequence alignment of cDNA subclone PP3#3 with mouse CYP4A14 by BLAST searching with the National Center --- p.374 / Chapter C29.3 --- Summary of sequence alignment of cDNA subclone PP3#3 with mouse CYP4A14 --- p.374 / Chapter C30.1 --- DNA sequence of cDNA subclone PP3#13 using Ml3 reverse primer and the sequence of 5'-ARP3 --- p.375 / Chapter C30.2 --- Sequence alignment of cDNA subclone PP3#13 with mouse CYP4A14 by BLAST searching with the National Center for Biotechnology Information database --- p.375 / Chapter C30.3 --- Summary of sequence alignment of cDNA subclone PP3#13 with mouse CYP4A14 --- p.375 / Chapter C31.1 --- DNA sequence of cDNA subclone QQ4#14 using Ml3 forward -20 primer and the sequence of 5´ة-ARP2 --- p.376 / Chapter C31.2 --- Sequence alignment of cDNA subclone QQ4#14 with mouse MCAD by BLAST searching with the National Center for Biotechnology Information database --- p.376 / Chapter C31.3 --- Summary of sequence alignment of cDNA subclone QQ4#14 with mouse MCAD --- p.376 / Chapter C32.1 --- DNA sequence of cDNA subclone QQ4#15 using Ml3 reverse primer and the sequence of 5´ة-ARP2 --- p.377 / Chapter C32.2 --- Sequence alignment of cDNA subclone QQ4#15 with mouse MCAD by BLAST searching with the National Center for Biotechnology Information database --- p.377 / Chapter C32.3 --- Summary of sequence alignment of cDNA subclone QQ4#15 with mouse MCAD --- p.377 / Chapter C33.1 --- DNA sequence of cDNA subclone QQ5#14 using Ml3 forward -20 primer and the sequence of 3'-AP3 --- p.377 / Chapter C33.2 --- Sequence alignment of cDNA subclone QQ5#14 with mouse MUP II by BLAST searching with the National Center for Biotechnology Information database --- p.378 / Chapter C33.3 --- Summary of sequence alignment of cDNA subclone QQ5#14 with mouse MUP II --- p.378 / Chapter C34.1 --- DNA sequence of cDNA subclone QQ5#17 using Ml3 reverse primer and the sequence of 5'-ARP2 --- p.378 / Chapter C34.2 --- Sequence alignment of cDNA subclone QQ5#17 with mouse MUP II by BLAST searching with the National Center for Biotechnology Information database --- p.379 / Chapter C34.3 --- Summary of sequence alignment of cDNA subclone QQ5#17 with mouse MUP II --- p.379 / Chapter C35.1 --- DNA sequence of cDNA subclone QQ6#13 using Ml3 forward -20 primer and the sequence of 3'-AP3 --- p.380 / Chapter C35.2 --- Sequence alignment of cDNA subclone QQ6#13 with mouse MUP II by BLAST searching with the National Center for Biotechnology Information database --- p.380 / Chapter C35.3 --- Summary of sequence alignment of cDNA subclone QQ6#13 with mouse MUP II --- p.380 / Chapter C36.1 --- DNA sequence of cDNA subclone QQ6#14 using M13 reverse primer and the 5'-ARP2 --- p.381 / Chapter C36.2 --- Sequence alignment of cDNA subclone QQ6#14 with mouse MUP II by BLAST searching with the National Center for Biotechnology Information database --- p.381 / Chapter C36.3 --- Summary of sequence alignment of cDNA subclone QQ6#14 with mouse MUP II --- p.381 / Chapter C37.1 --- DNA sequence of cDNA subclone TT4#5 using Ml3 forward -20 primer and the sequence of 3´ة-AP5 --- p.382 / Chapter C37.2 --- Sequence alignment of cDNA subclone TT4#5 with mouse MUP II by BLAST searching with the National Center for Biotechnology Information database --- p.382 / Chapter C37.3 --- Summary of sequence alignment of cDNA subclone TT4#5 with mouse MUP II --- p.382 / Chapter C38.1 --- DNA sequence of cDNA subclone TT4#8 using Ml3 forward -20 primer and the sequence of 5´ة-ARP2 --- p.383 / Chapter C38.2 --- Sequence alignment of cDNA subclone TT4#8 with mouse MUP II by BLAST searching with the National Center for Biotechnology Information database --- p.383 / Chapter C38.3 --- Summary of sequence alignment of cDNA subclone TT4#8 with mouse MUP II --- p.383 / Chapter C39.1 --- DNA sequence of cDNA subclone TT4#9 using Ml3 reverse primer and the sequence of 3'-AP5 --- p.384 / Chapter C39.2 --- Sequence alignment of cDNA subclone TT4#9 with mouse MUP II by BLAST searching with the National Center for Biotechnology Information database --- p.384 / Chapter C39.3 --- Summary of sequence alignment of cDNA subclone TT4#9 with mouse MUP II --- p.384 / Chapter C40.1 --- DNA sequence of cDNA subclone TT5#1 using Ml3 forward -20 primer and the sequence of 5'-ARP2 --- p.385 / Chapter C40.2 --- Sequence alignment of cDNA subclone TT5#1 with mouse MUP II by BLAST searching with the National Center for Biotechnology Information database --- p.385 / Chapter C40.3 --- Summary of sequence alignment of cDNA subclone TT5#1 with mouse MUP II --- p.385 / Chapter C41.1 --- DNA sequence of cDNA subclone TT5#9 using Ml3 reverse primer and the sequence of 3´ة-AP5 --- p.386 / Chapter C41.2 --- Sequence alignment of cDNA subclone TT5#9 with mouse MUP II by BLAST searching with the National Center for Biotechnology Information database --- p.386 / Chapter C41.3 --- Summary of sequence alignment of cDNA subclone TT5#9 with mouse MUP II --- p.386 / Chapter C42.1 --- DNA sequence of cDNA subclone UU1#1 using Ml3 forward -20 primer and the sequence of 3´ة-AP2 --- p.387 / Chapter C42.2 --- Sequence alignment of cDNA subclone UU1#1 with mouse CYP4A10 by BLAST searching with the National Center for Biotechnology Information database --- p.387 / Chapter C42.3 --- Summary of sequence alignment of cDNA subclone UU1#1 with mouse CYP4A10 --- p.387 / Chapter C43.1 --- DNA sequence of cDNA subclone UU1#4 using Ml3 reverse primer and the sequence of 5'-ARP12 --- p.388 / Chapter C43.2 --- Sequence alignment of cDNA subclone UU1#4 with mouse CYP4A10 by BLAST searching with the National Center for Biotechnology Information database --- p.388 / Chapter C43.3 --- Summary of sequence alignment of cDNA subclone UU1#4 with mouse CYP4A10 --- p.388 / Chapter C44.1 --- DNA sequence of cDNA subclone VV5#2 using Ml3 forward -20 primer and the sequence of 3´ة-AP2 --- p.389 / Chapter C44.2 --- Sequence alignment of cDNA subclone VV5#2 with mouse CYP4A10 by BLAST searching with the National Center for Biotechnology Information database --- p.389 / Chapter C44.3 --- Summary of sequence alignment of cDNA subclone VV5#2 with mouse CYP4A10 --- p.389 / Chapter C45.1 --- DNA sequence of cDNA subclone VV5#3 using Ml3 forward -20 primer and the sequence of 3'-AP2 --- p.390 / Chapter C45.2 --- Sequence alignment of cDNA subclone VV5#3 with mouse CYP4A10 by BLAST searching with the National Center for Biotechnology Information database --- p.390 / Chapter C45.3 --- Summary of sequence alignment of cDNA subclone VV5#3 with mouse CYP4A10 --- p.390 / Chapter C46.1 --- DNA sequence of cDNA subclone VV5#4 using Ml3 reverse primer and the sequence of 5'-ARP18 --- p.391 / Chapter C46.2 --- Sequence alignment of cDNA subclone VV5#4 with mouse CYP4A10 by BLAST searching with the National Center for Biotechnology Information database --- p.391 / Chapter C46.3 --- Summary of sequence alignment of cDNA subclone VV5#4 with mouse CYP4A10 --- p.391 / Chapter C47.1 --- DNA sequence of cDNA subclone VV8#5 using M13 forward -20 primer and the sequence of 5'-ARP18 --- p.392 / Chapter C47.2 --- Sequence alignment of cDNA subclone VV8#5 with mouse HMG-CoAS by BLAST searching with the National Center for Biotechnology Information database --- p.392 / Chapter C47.3 --- Summary of sequence alignment of cDNA subclone VV8#5 with mouse HMG-CoAS --- p.392 / Chapter C48.1 --- DNA sequence of cDNA subclone VV8#6 using Ml 3 reverse primer and the sequence of 3'-AP2 --- p.393 / Chapter C48.2 --- Sequence alignment of cDNA subclone VV8#6 with mouse HMG-CoAS by BLAST searching with the National Center for Biotechnology Information database --- p.393 / Chapter C48.3 --- Summary of sequence alignment of cDNA subclone VV8#6 with mouse HMG-CoAS --- p.393 / Chapter C49.1 --- DNA sequence of cDNA subclone WW1#3 using Ml 3 forward -20 primer and the sequence of 3'-AP2 --- p.394 / Chapter C49.2 --- Sequence alignment of cDNA subclone WW1#3 with mouse Spil-3 by BLAST searching with the National Center for Biotechnology Information database --- p.394 / Chapter C49.3 --- Summary of sequence alignment of cDNA subclone WW1#3 with mouse Spil-3 --- p.394 / Chapter C50.1 --- DNA sequence of cDNA subclone WW1#4 using Ml3 forward -20 primer and the sequence of 3'-AP2 --- p.395 / Chapter C50.2 --- Sequence alignment of cDNA subclone WW1#4 with mouse Spil-3 by BLAST searching with the National Center for Biotechnology Information database --- p.395 / Chapter C50.3 --- Summary of sequence alignment of cDNA subclone WW1#4 with mouse Spil-3 --- p.395 / Chapter C51.1 --- DNA sequence of cDNA subclone WW1#7 using M13 reverse primer and the sequence of 5´ة-ARP19 --- p.396 / Chapter C51.2 --- Sequence alignment of cDNA subclone WW1#7 with mouse Spil-3 by BLAST searching with the National Center for Biotechnology Information database --- p.396 / Chapter C51.3 --- Summary of sequence alignment of cDNA subclone WW1#7 ith mouse Spil-3 --- p.396 / Chapter C52.1 --- DNA sequence of cDNA subclone XX1#2 using Ml3 forward 20 primer and the sequence of 3´ة-APl0 --- p.397 / Chapter C52.2 --- Sequence alignment of cDNA subclone XX1#2 with mouse CYP4A10 by BLAST searching with the National Center for Biotechnology Information database --- p.397 / Chapter C52.3 --- Summary of sequence alignment of cDNA subclone XX1#2 with mouse CYP4A10 --- p.397 / Chapter C53.1 --- DNA sequence of cDNA subclone XX1#3 using M13 forward -20 primer and the sequence of 3´ة-APl0 D205 --- p.398 / Chapter C53.2 --- Sequence alignment of cDNA subclone XX1#3 with mouse CYP4A10 by BLAST searching with the National Center for Biotechnology Information database --- p.398 / Chapter C53.3 --- Summary of sequence alignment of cDNA subclone XX1#3 with mouse CYP4A10 --- p.398 / Chapter C54.1 --- DNA sequence of cDNA subclone XX1#9 using Ml3 forward -20 primer and the sequence of 5'-ARP12 --- p.399 / Chapter C54.2 --- Sequence alignment of cDNA subclone XX1#9 with mouse CYP4A10 by BLAST searching with the National Center for Biotechnology Information database --- p.399 / Chapter C54.3 --- Summary of sequence alignment of cDNA subclone XX1#9 with mouse CYP4A10 --- p.399

Thesis (Ph.D.)--McMaster University (Canada), 1995. / Source: Dissertation Abstracts International, Volume: 56-12, Section: B, page: 6733. Adviser: R. A. Rachubinski.

No description available.

The oxidation status of a cell plays a crucial role in aging. As cells get aged, their redox state gets increased. Pex5p is a peroxisomal recycling receptor which binds to newly synthesized cargo proteins in the cytosol and imports them across the peroxisomal membrane. During this transport event, Pex5p gets monoubiquitinated at a conserved cysteine (C11) residue. This C11 is very essential for the recycling of Pex5p from the peroxisomal membrane to back into the cytosol. If the cysteine is replaced by serine, Pex5p does not get recycled back to the cytosol and accumulates on the peroxisomal membrane. In the present study, we have investigated whether the C11 in Pex5p could act as a redox switch. We measured the redox state of the cytosol and the peroxisomal matrix as well as the subcellular localization of catalase in aging cells. We found that an increase in the redox state of peroxisomes (in WT) leads to an increase in the redox state of the cytosol, which ultimately results in the impairment of PTS1 import. Interestingly, in the C11K condition, we did not see an impairment of PTS1 import. These observations support our hypothesis that C11 may act as a redox switch. We also performed some challenging experiments with H2O2. The results of these experiments show that a) import of catalase into peroxisomes sensitizes the cytosol and b) catalase overexpression does have a protective effect against oxidative stress caused by H2O2. In summary the results of our experiments support our hypothesis. However, further evaluation is needed to reveal the precise role of C11 in Pex5p function during cellular aging.

Thesis (Ph. D. in Cell Biology)--Vanderbilt University, May 2004. / Title from PDF title screen. Includes bibliographical references.