Nucleolin stabilizes Bcl-X L messenger RNA in response to UVA irradiation.
Our laboratory has previously reported that UVA irradiation can increase the expression of Bcl-X(L), an antiapoptotic molecule, by stabilizing its mRNA in cultured immortalized human keratinocytes. To understand the mechanism by which the Bcl-X(L) message is stabilized, we used a synthetic Bcl-X(L) 3'-untranslated region (UTR) to capture RNA-binding proteins. Nucleolin was identified as one of the binding proteins as determined by tandem mass spectrometry coupled to liquid chromatography analysis. Further study showed that nucleolin specifically recognized the AU-rich elements (AUUUA) in the 3'-UTR of the Bcl-X(L) mRNA and could stabilize the mRNA in vitro. Furthermore, overexpression of nucleolin stabilizes the Bcl-X(L) mRNA in HeLa cells, whereas reducing nucleolin by small interfering RNA shortens the Bcl-X(L) mRNA half-life. Interestingly, nucleolin physically interacted with polyadenylate [poly(A)]-binding protein through it RGG motifs. Its stabilizing effect on the Bcl-X(L) mRNA was dependent upon the presence of poly(A) tail. Based on these data, we propose a model in which nucleolin protects the Bcl-X(L) mRNA from nuclease degradation by enhancing the stability of the ribonucleoprotein loop structure.
Zhang
Jack
J
Arizona Cancer Center, University of Arizona, Tuczon, Arizona 85724, USA.
Tsaprailis
George
G
Bowden
G Tim
GT
D020413:3' Untranslated Regions; D006367:HeLa Cells; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism / Q000528:radiation effects; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism*; D039102:Poly(A)-Binding Proteins / Q000378:metabolism; D011485:Protein Binding; D012333:RNA, Messenger / Q000378:metabolism* / Q000528:radiation effects; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D014466:Ultraviolet Rays; D051020:bcl-X Protein / Q000235:genetics*; D000097796:Nucleolin
D020413:3' Untranslated Regions; D010750:Phosphoproteins; D039102:Poly(A)-Binding Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D051020:bcl-X Protein; D000097796:Nucleolin; C494811:BCL2L1 protein, human