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PMID: 3368312 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Initiator role of double stranded DNA in terminal transferase catalyzed polymerization reactions.

Nucleic acids research ·Vol. 16 ·No. 7 ·1988-04-11 ·Pages 2943-57

Robbins DJ, Coleman MS

Abstract

Binding of the 58 kDa monomer and 44 kDa alpha beta dimer forms of terminal deoxynucleotidyl transferase to double stranded DNA was demonstrated by gel retardation and tryptophan fluorescence quenching. The dissociation constants and cooperativity parameters were similar to those that have been determined for binding of these two forms of terminal transferase to single stranded DNA. However, the double stranded DNA binding site size of 10 nucleotides was half the size expected. The efficacy of blunt ended DNA as an initiator in the polymerization reaction catalyzed by terminal transferase was demonstrated by radiometric assays and product analyses on agarose gels. The initial reaction kinetics indicated that dGTP but not dATP was added efficiently to a blunt double stranded DNA 3' end. These results are correlated with current models for in vivo terminal transferase function.

MeSH Terms
Base Sequence DNA/metabolism,physiology DNA Nucleotidylexotransferase/metabolism,physiology DNA Nucleotidyltransferases/metabolism DNA, Single-Stranded/physiology DNA-Directed DNA Polymerase/metabolism Electrophoresis, Agar Gel Kinetics Peptide Chain Initiation, Translational Proteins/pharmacology Spectrometry, Fluorescence
Chemicals
DNA, Single-Stranded Proteins DNA DNA Nucleotidyltransferases DNA Nucleotidylexotransferase DNA-Directed DNA Polymerase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Robbins D J
Department of Biochemistry, University of Kentucky Medical Center, Lexington 40536-0084.
Coleman M S
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/16.7.2943
Published
1988-04-11
Pages
2943-57
Language
English
Region
England
NLM ID
0411011
PMCID
PMC336443
Subset
IM
Grants
NCI NIH HHS · CA19492 · United States
NCI NIH HHS · CA26391 · United States
External Links
PubMed source DOI: 10.1093/nar/16.7.2943 PMC Full Text View journal details
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