Deletion mutational analysis of BMRP, a pro-apoptotic protein that binds to Bcl-2.
Bcl-2 is an anti-apoptotic member of the Bcl-2 family of proteins that protects cells from apoptosis induced by a large variety of stimuli. The protein BMRP (MRPL41) was identified as a Bcl-2 binding partner and shown to have pro-apoptotic activity. We have performed deletion mutational analyses to identify the domain(s) of Bcl-2 and BMRP that are involved in the Bcl-2/BMRP interaction, and the region(s) of BMRP that mediate its pro-apoptotic activity. The results of these studies indicate that both the BH4 domain of Bcl-2 and its central region encompassing its BH1, BH2, and BH3 domains are required for its interaction with BMRP. The loop region and the transmembrane domain of Bcl-2 were found to be dispensable for this interaction. The Bcl-2 deletion mutants that do not interact with BMRP were previously shown to be functionally inactive. Deletion analyses of the BMRP protein delimited the region of BMRP needed for its interaction with Bcl-2 to the amino-terminal two-thirds of the protein (amino acid residues 1-92). Further deletions at either end of the BMRP(1-92) truncated protein resulted in lack of binding to Bcl-2. Functional studies performed with BMRP deletion mutants suggest that the cell death-inducing domains of the protein reside mainly within its amino-terminal two-thirds. The region of BMRP required for the interaction with Bcl-2 is very relevant for the cell death-inducing activity of the protein, suggesting that one possible mechanism by which BMRP induces cell death is by binding to and blocking the anti-apoptotic activity of Bcl-2.
Department of Chemistry, Texas A&M University-Kingsville, 700 University Blvd., Kingsville, TX 78363-8202, USA.
Srinivas
S
Malladi
Kishore V L
KV
Parsa
Deepthi
D
Bhupathi
María A
MA
Rodríguez-González
Juan A
JA
Conde
Pallavi
P
Anumula
Hannah E
HE
Romo
Cheryl J
CJ
Claunch
Rafael P
RP
Ballestero
Maribel
M
González-García
D000818:Animals; D017209:Apoptosis; D001483:Base Sequence; D015153:Blotting, Western; D017931:DNA Primers; D006801:Humans; D051379:Mice; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D041681:NIH 3T3 Cells; D011485:Protein Binding; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000235:genetics / Q000378:metabolism*; D017384:Sequence Deletion; D020798:Two-Hybrid System Techniques
D017931:DNA Primers; C502654:MRPL41 protein, human; D024101:Mitochondrial Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012269:Ribosomal Proteins
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