Thesis (Ph. D.)--University of Washington, 2000. / Vita. Includes bibliographical references (leaves 140-160).

A biochemical approach was used to detect the appearance of the heavy chain of skeletal myosin (HCSM) and myosin mRNA during the early development of Xenopus laevis embryos. An antibody against the HCSM of adult X. laevis muscles was biochemically characterized and shown to be monospecific. This anti-myosin antibody reacted with embryonic polysomes synthesizing myosin and with tadpole tail myosin. This indicates that the myosins of adult muscles, early embryonic muscles and tadpole tails are sufficiently homologous to share some antigenic determinants. Polysomes from various stages of X. laevis embryogenesis were reacted with the anti-myosin antibody. Analysis of these reactions showed that myosin synthesis begins in stage 20 embryos, in which about 7 somites have segregated. The RNA from stage 12, stage 16/17 and stage 20 embryos was then analyzed for the presence of the heavy chain myosin mRNA in order to determine whether the synthesis of myosin is under translational or transcriptional control. Total RNA preparations from staged embryos were fractionated on oligo(dT)-cellulose columns and fractions that did and did not bind were translated in a wheat germ cell-free protein synthesizing system. The translational products were precipitated with the anti-myosin antibody and characterized biochemically. Myosin mRNA was detected by this method in stage 16/17 embryos. We conclude that somite segregation results in the appearance of new myosin mRNA molecules in X. laevis embryos. It seems likely, by all the evidence considered, that a large pool of untranslated myosin mRNA molecules is not responsible for muscle myosin synthesis. Therefore, the synthesis of certain proteins in early development is under transcriptional control. / Science, Faculty of / Zoology, Department of / Unknown

Thesis (Ph. D.)--Case Western Reserve University, 2008. / [School of Medicine] Department of Pharmacology. Includes bibliographical references.

Thesis (M.S.)--West Virginia University, 2010. / Title from document title page. Document formatted into pages; contains viii, 37 p. : ill. (some col.). Includes abstract. Includes bibliographical references (p. 33-37).

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Thesis (Ph. D.)--University of Missouri-Columbia, 2006. / The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Vita. "August 2006" Includes bibliographical references.

The class I myosins are an ubiquitous family of non-filamentous, single-headed actin-binding motor proteins. The objective of this study was to identify the light chain composition of the short-tailed Dictyostelium class I myosins, MyoIA and MyoIE. Flag-tagged MyoIA head-neck and MyoIE head-neck constructs were generated and expressed in Dicyostelium discoidem. The MyoIA and MyoIE head-neck constructs both co-purified with a 17-kDa protein that reacted with an anti-calmodulin antibody and exhibited a mobility shift on SDS gels in the presence of calcium. Mass spectrometry analysis confirmed that the light chain bound to MyoIA and MyoIE was calmodulin. The finding that the short-tailed class I Dictyostelium myosins use the generic calcium-binding protein calmodulin as a light chain contrasts with previous work showing that the long-tailed Dictyostelium class I myosins MyoIB, MyoIC, and MyoID each bind a unique, specialized light chain called MlcB, MlcC, and MlcD, respectively. Despite having a calmodulin light chain, calcium did not affect the actin-activated Mg-ATPase activities of MyoIA or MyoIE. The p21-activated kinases (PAKs) are serine-threonine protein kinases that are activated by the small GTPases Cdc42 and Rac. PAKs phosphorylate a site in the motor domain of Dictyostelium class I myosins that is required for myosin activity. Studies were carried out to determine whether Dictyostelium RacB, which is known to bind to and activate Dictyostelium PAKs, promotes the phosphorylation of MyoID in vivo. A vector that expresses a constitutively active RacB under the control of a doxycycline-inducible promoter was created and transformed into Dictyostelium cells. Immunostaining demonstrated that the constitutively active RacB increased actin filament formation in AX3 cells by ~3-fold but by only ~1.5-fold in PakB-null cells. A rabbit polyclonal antibody against the MyoID tail was made. An anti-phospho antibody raised against a phosphorylated peptide corresponding to the MyoID TEDS site was tested and found to specifically recognize purified phosphorylation MyoIA and MyoID. The anti-phospho antibody did not detect phosphorylated MyoIA or MyoID in crude Dictyostelium cell extracts or in immunoprecipitates prepared using the anti-MyoID antibody. Further work is needed to improve the specificity of the anti-phospho MyoID antibody. / Thesis (Master, Biochemistry) -- Queen's University, 2009-09-24 19:51:55.032

Thesis (Ph. D.)--University of Washington, 2007. / Vita. Includes bibliographical references (p. 150-159).

Thesis (Ph. D.)--University of Washington, 1997. / Vita. Includes bibliographical references (leaves [67]-75).