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PMID: 15070731 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Mammalian GLD-2 homologs are poly(A) polymerases.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 101 ·No. 13 ·2004-03-30 ·Pages 4407-12

Kwak JE, Wang L, Ballantyne S, Kimble J, Wickens M

Abstract

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.

MeSH Terms
Animals Humans Kinetics Mice Models, Molecular Nucleotidyltransferases/genetics,metabolism Phylogeny Plasmids Poly A/metabolism Polynucleotide Adenylyltransferase/chemistry,genetics,metabolism Protein Biosynthesis Protein Conformation RNA, Messenger/genetics Recombinant Fusion Proteins/chemistry,metabolism Transcription, Genetic
Chemicals
RNA, Messenger Recombinant Fusion Proteins Poly A Nucleotidyltransferases Polynucleotide Adenylyltransferase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Kwak Jae Eun
Department of Biochemistry, 433 Babcock Drive, University of Wisconsin, Madison, WI 53706-1544, USA.
Wang Liaoteng
Ballantyne Scott
Kimble Judith
Wickens Marvin
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0400779101
Published
2004-03-30
Pages
4407-12
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC384760
Subset
IM
Grants
NIGMS NIH HHS · R01 GM031892 · United States
NIGMS NIH HHS · R37 GM031892 · United States
NIGMS NIH HHS · GM31892 · United States
External Links
PubMed source DOI: 10.1073/pnas.0400779101 PMC Full Text View journal details
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