Purification and characterization of TnsC, a Tn7 transposition protein that binds ATP and DNA

Pascal Gamas, Nancy L Craig

Research output: Contribution to journalArticle

Abstract

The bacterial transposon Tn7 encodes five transposition genes tnsABCDE. We report a simple and rapid procedure for the purification of TnsC protein. We show that purified TnsC is active in and required for Tn7 transposition in a cell-free recombination system. This finding demonstrates that TnsC participates directly in Tn7 transposition and explains the requirement for tnsC function in Tn7 transposition. We have found that TnsC binds adenine nucleotides and is thus a likely site of action of the essential ATP cofactor in Tn7 transposition. We also report that TnsC binds non-specifically to DNA in the presence of ATP or the generally non-hydrolyzable analogues AMP-PNP and ATP-γ-S, and that TnsC displays little affinity for DNA in the presence of ADP. We speculate that TnsC plays a central role in the selection of target DNA during Tn7 transposition.

Original languageEnglish (US)
Pages (from-to)2525-2532
Number of pages8
JournalNucleic Acids Research
Volume20
Issue number10
StatePublished - May 25 1992

Fingerprint

Transposition
Adenosinetriphosphate
Purification
DNA
Adenosine Triphosphate
Proteins
Protein
Adenylyl Imidodiphosphate
Administrative data processing
Adenine Nucleotides
Cell-Free System
Nucleotides
Adenosine Diphosphate
Genetic Recombination
Genes
Cofactor
Recombination
Affine transformation
Gene
Likely

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

Purification and characterization of TnsC, a Tn7 transposition protein that binds ATP and DNA. / Gamas, Pascal; Craig, Nancy L.

In: Nucleic Acids Research, Vol. 20, No. 10, 25.05.1992, p. 2525-2532.

Research output: Contribution to journalArticle

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