SNAMA is a 142 kDa Drosophila melanogaster protein, which consists of the
uncharacterized conserved domain with no name (DWNN), zinc and RING finger-like
motifs. The primary structure of SNAMA suggests that it might play an important role in
cell cycle regulation and apoptosis. Previous studies revealed that homozygous SNAMA
mutants underwent ectopic apoptosis which resulted in recessive lethality. SNAMA
orthologues such P2P-R, PACT and RBBP6 are involved in cell cycle regulation,
whereas Mpe1 is involved in mRNA processing. The aim of this study was to map out
the role of SNAMA by isolating proteins which interact with it. DWNN was inserted
into pGEX6P-2, phylexzeo plasmid (bait) and the Drosophila 0-12 hours cDNA library
inserted in pJG4-5 (prey). The bait and the prey plasmid were used to transform
appropriate yeast cells to probe for interacting proteins in yeast two hybrid assays,
whereas the pGEX6P-2 was used for heterologous overexpression of DWNN in E. coli.
Immunoprecipitation assays were also carried out with the crude protein extract from
embryos, adult wild type, SNAMA mutant flies and the overexpressed protein using
antibodies against SNAMA, Drosophila p53 Human DWNN and GST. The hybrid assay
did not produce any interactors. Some of the proteins obtained from the
immunoprecipitations were isolated and sequenced. The proteins identified were hsp82,
Hsp70 and CG2985-PA. Data obtained from the immunoprecipitations suggest that
SNAMA like Dmp53 might be involved in cell cycle regulation.
| Identifer | oai:union.ndltd.org:netd.ac.za/oai:union.ndltd.org:wits/oai:wiredspace.wits.ac.za:10539/5784 |
| Date | 22 October 2008 |
| Creators | Rakgotho, Patrick Mpho |
| Source Sets | South African National ETD Portal |
| Language | English |
| Detected Language | English |
| Type | Thesis |
| Format | application/pdf |
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