Mammalian GLD-2 homologs are poly(A) polymerases.
GLD-2 is a cytoplasmic poly(A) polymerase present in the Caenorhabditis elegans germ line and embryo. It is a divergent member of the DNA polymerase beta nucleotidyl transferase superfamily, which includes CCA-adding enzymes, DNA polymerases and eukaryotic nuclear poly(A) polymerases. The polyadenylation activity of GLD-2 is stimulated by physical interaction with an RNA binding protein, GLD-3. To test whether GLD-3 might stimulate GLD-2 by recruiting it to RNA, we tethered C. elegans GLD-2 to mRNAs in Xenopus oocytes by using MS2 coat protein. Tethered GLD-2 adds poly(A) and stimulates translation of the mRNA, demonstrating that recruitment is sufficient to stimulate polyadenylation activity. We use the same tethered assay to identify human and mouse poly(A) polymerases related to GLD-2. This may provide entrees to previously uncharacterized modes of polyadenylation in mammalian cells.
Kwak
Jae Eun
JE
Department of Biochemistry, 433 Babcock Drive, University of Wisconsin, Madison, WI 53706-1544, USA.
Wang
Liaoteng
L
Ballantyne
Scott
S
Kimble
Judith
J
Wickens
Marvin
M
D000818:Animals; D006801:Humans; D007700:Kinetics; D051379:Mice; D008958:Models, Molecular; D009713:Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D010802:Phylogeny; D010957:Plasmids; D011061:Poly A / Q000378:metabolism; D011062:Polynucleotide Adenylyltransferase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D014176:Protein Biosynthesis; D011487:Protein Conformation; D012333:RNA, Messenger / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D014158:Transcription, Genetic
D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D011061:Poly A; D009713:Nucleotidyltransferases; D011062:Polynucleotide Adenylyltransferase