title: Annexin A2 and PSF proteins interact with p53 IRES and regulate translation of p53 mRNA.
abstract: p53 mRNA has been shown to be translated into two isoforms, full-length p53 (FL-p53) and a truncated isoform ΔN-p53, which modulates the functions of FL-p53 and also has independent functions. Previously, we have shown that translation of p53 and ΔN-p53 can be initiated at Internal Ribosome Entry Sites (IRES). These two IRESs were shown to regulate the translation of p53 and ΔN-p53 in a distinct cell-cycle phase-dependent manner. Earlier observations from our laboratory also suggest that the structural integrity of the p53 RNA is critical for IRES function and is compromised by mutations that affect the structure as well as RNA protein interactions. In the current study, using RNA affinity approach we have identified Annexin A2 and PTB associated Splicing Factor (PSF/SFPQ) as novel ITAFs for p53 IRESs. We have showed that the purified Annexin A2 and PSF proteins specifically bind to p53 IRES elements. Interestingly, in the presence of calcium ions Annexin A2 showed increased binding with p53 IRES. Immunopulldown experiments suggest that these two proteins associate with p53 mRNA ex vivo as well. Partial knockdown of Annexin A2 and PSF showed decrease in p53 IRES activity and reduced levels of both the p53 isoforms. More importantly the interplay between Annexin A2, PSF and PTB proteins for binding to p53mRNA appears to play a crucial role in IRES function. Taken together, our observations suggest pivotal role of two new trans-acting factors in regulating the p53-IRES function, which in turn influences the synthesis of p53 isoforms.
authors.affiliation: Department of Microbiology and Cell Biology; Indian Institute of Science; Bangalore, India.
authors.forename: Arandkar
authors.initials: A
authors.lastname: Sharathchandra
authors.forename: Ridhima
authors.initials: R
authors.lastname: Lal
authors.forename: Debjit
authors.initials: D
authors.lastname: Khan
authors.forename: Saumitra
authors.initials: S
authors.lastname: Das
mesh_terms: D017306:Annexin A2 / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D045744:Cell Line, Tumor; D059865:Endoplasmic Reticulum Stress; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D006801:Humans; D000072670:PTB-Associated Splicing Factor; D011485:Protein Binding; D014176:Protein Biosynthesis; D025941:Protein Interaction Mapping; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*
chemical_list: C472822:ANXA2 protein, human; D017306:Annexin A2; D000072670:PTB-Associated Splicing Factor; D020033:Protein Isoforms; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D002118:Calcium
keywords: IRES; IRES trans-acting factors; PSF; internal initiation of translation Annexin A2; p53mRNA