Ornithine-$ delta$-biaminotransferase (OAT) is a mitochondrial matrix a enzyme which catalyzes the reversible conversion of ornithine to glutamic semialdehyde. In the retinoblastoma cell line Y79, the 5$ sp prime$ UTR of the OAT mRNA is alternatively-spliced. Previous work has shown that the two alternatively-spliced 5$ sp prime$ UTR are translated with different efficiencies. / The alternatively-spliced 5$ sp prime$ UTRs were isolated and subcloned. Expression vectors were constructed containing each of the OAT 5$ sp prime$ UTRs and the human growth hormone as a reporter gene. Transient transfection expression analysis in COS-1 cells confirmed that the 5$ sp prime$ UTRs conferred differential expression on the reporter gene. The shorter 5$ sp prime$ UTR was expressed at a level 3-fold higher than that of the longer 5$ sp prime$ UTR. / RNA mobility shift assays were performed to determine whether a transacting factor was interacting with one of the alternatively-spliced 5$ sp prime$ UTRs. A binding activity was detected that bound to the longer, exon 2-containing 5$ sp prime$ UTR with a significantly higher affinity than the shorter 5$ sp prime$ UTR. Competition studies demonstrated that this binding activity was specific for the longer 5$ sp prime$ UTR.
Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.23283 |
Date | January 1995 |
Creators | MacDonald, Heather R. (Heather Ruth) |
Publisher | McGill University |
Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
Language | English |
Detected Language | English |
Type | Electronic Thesis or Dissertation |
Format | application/pdf |
Coverage | Master of Science (Department of Biology.) |
Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
Relation | alephsysno: 001481980, proquestno: MM08025, Theses scanned by UMI/ProQuest. |
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