The La autoantigen contains a dimerization domain that is essential for enhancing translation.
The La autoantigen is an RNA-binding protein that is involved in initiation and termination of RNA polymerase III transcription. It also binds several viral RNAs, including those of poliovirus and human immunodeficiency virus (HIV). Binding of the La protein to these RNAs enhances their translation in vitro (K. Meerovitch, Y.V. Svitkin, H.S. Lee, F. Lejbkowicz, D.J. Kenan, E.K.L. Chan, V.L. Agol, J.D. Keene, and N. Sonenberg, J. Virol. 67:3798-3807, 1993, and Y.V. Svitkin, A. Pause, and N. Sonenberg, J. Virol. 68:7001-7007, 1994). Here, a functional domain in the carboxy-terminal half of La that is distinct from the RNA-binding domain is described. Deletion of this domain abrogated the ability of La protein to enhance translation of poliovirus RNA and a hybrid HIV trans-activation-response element-chloramphenicol acetyltransferase mRNA. Far-Western assays indicated that the La protein homodimerized in vitro, and the C-terminal deletions that caused a loss of activity in translation also abrogated the dimerization signal. Gel filtration chromatography of recombinant La protein confirmed that La protein exists as a dimer under native conditions. Addition of the purified dimerization domain resulted in a loss of translation stimulatory activity of La protein in cell-free-translation reactions.
Craig
A W
AW
Department of Biochemistry, McGill University, Montreal, Quebec, Canada.
Svitkin
Y V
YV
Lee
H S
HS
Belsham
G J
GJ
Sonenberg
N
N
D001324:Autoantigens / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D019281:Dimerization; D015497:HIV-1 / Q000235:genetics; D006801:Humans; D008970:Molecular Weight; D017955:Poliovirus / Q000235:genetics; D014176:Protein Biosynthesis / Q000187:drug effects*; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000302:isolation & purification; D012261:Ribonucleoproteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology*; D017384:Sequence Deletion; D000097585:SS-B Antigen
D001324:Autoantigens; D015500:Chloramphenicol O-Acetyltransferase; D012313:RNA; D012333:RNA, Messenger; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D012261:Ribonucleoproteins; D000097585:SS-B Antigen; C039919:RNA, recombinant