Identification of proteins that interact with DWNN domain of SNAMA a member of a novel protein superfamily
Year: 2008
Type: dissertation
Abstract: SNAMA is a 142 kDa Drosophila melanogaster protein, which consists of the \nuncharacterized conserved domain with no name (DWNN), zinc and RING finger-like \nmotifs. The primary structure of SNAMA suggests that it might play an important role in \ncell cycle regulation and apoptosis. Previous studies revealed that homozygous SNAMA \nmutants underwent ectopic apoptosis which resulted in recessive lethality. SNAMA \northologues such P2P-R, PACT and RBBP6 are involved in cell cycle regulation, \nwhereas Mpe1 is involved in mRNA processing. The aim of this study was to map out \nthe role of SNAMA by isolating proteins which interact with it. DWNN was inserted \ninto pGEX6P-2, phylexzeo plasmid (bait) and the Drosophila 0-12 hours cDNA library \ninserted in pJG4-5 (prey). The bait and the prey plasmid were used to transform \nappropriate yeast cells to probe for interacting proteins in yeast two hybrid assays, \nwhereas the pGEX6P-2 was used for heterologous overexpression of DWNN in E. coli. \nImmunoprecipitation assays were also carried out with the crude protein extract from \nembryos, adult wild type, SNAMA mutant flies and the overexpressed protein using \nantibodies against SNAMA, Drosophila p53 Human DWNN and GST. The hybrid assay \ndid not produce any interactors. Some of the proteins obtained from the \nimmunoprecipitations were isolated and sequenced. The proteins identified were hsp82, \nHsp70 and CG2985-PA. Data obtained from the immunoprecipitations suggest that \nSNAMA like Dmp53 might be involved in cell cycle regulation.
Author: Patrick Mpho Rakgotho
Language: English
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: green
OpenAlex ID: W2252643291