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PMID: 1756732 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Isolation and expression of cDNA clones encoding mammalian poly(A) polymerase.

The EMBO journal ·Vol. 10 ·No. 13 ·1991-12-00 ·Pages 4251-7

Wahle E, Martin G, Schiltz E, Keller W

Abstract

cDNA clones encoding mammalian poly(A) polymerase were isolated with probes generated by the polymerase chain reaction based on amino acid sequences derived from the purified enzyme. A bovine cDNA clone was obtained encoding a protein of 82 kDa. Expression in Escherichia coli resulted in the appearance of a poly(A) polymerase activity that was dependent on the addition of the purified specificity factor CPF and the presence of the polyadenylation signal AAUAAA in the RNA substrate. The activity copurified with a polypeptide of the expected size. A second class of cDNAs encoded a polypeptide of 43 kDa which was closely related to the N-terminal half of the 82 kDa protein. Northern blots showed two mRNAs of 4.2 and 2.4 kb that probably correspond to the two classes of cDNAs, as well as a third band of 1.3 kb. The sequence of the N-terminal half of bovine poly(A) polymerase is 47% identical with the amino acid sequence of the corresponding part of yeast poly(A) polymerase. Homologies to other proteins are of uncertain significance.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Blotting, Northern Cattle Chromatography, Liquid Cloning, Molecular DNA/genetics,isolation & purification Electrophoresis, Polyacrylamide Gel Escherichia coli/genetics Gene Expression Genes, Bacterial HeLa Cells Humans Molecular Sequence Data Plasmids Polymerase Chain Reaction Polynucleotide Adenylyltransferase/genetics RNA, Messenger/metabolism
Chemicals
RNA, Messenger DNA Polynucleotide Adenylyltransferase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Wahle E
Universität Basel, Abteilung Zellbiologie, Switzerland.
Martin G
Schiltz E
Keller W
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1991-12-00
Pages
4251-7
Language
English
Region
England
NLM ID
8208664
PMCID
PMC453177
Subset
IM
Databases
GENBANK
M62513, M62514, M62515, S65080, S65083, S72609, S72755, X59372, X59373, X61585
External Links
PubMed source PMC Full Text View journal details
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