DNA sequence analysis of the transposon Tn3

Three genes and three sites involved in transposition of Tn3

Fred Heffron, Brian J. McCarthy, Hisako Ohtsubo, Eiichi Ohtsubo

Research output: Contribution to journalArticle

283 Citations (Scopus)

Abstract

The complete nucleotide sequence of the transposon Tn3 and of 20 mutations which affect its transposition are reported. The mutations, generated in vitro by random insertion of synthetic restriction sites, proved to contain small duplications or deletions immediately adjacent to the new restriction site. By determining the phenotype and DNA sequence of these mutations we were able to generate an overlapping phenotypic and nucleotide map. This 4957 bp transposon encodes three polypeptides which account for all but 350 bp of its total coding capacity. These proteins are the transposase, a high molecular weight polypeptide (1015 amino acids) encoded by the tnpA gene; the Tn3-specific repressor, a low molecular weight polypeptide (185 amino acids) encoded by the tnpR gene; and the 286 amino acid β-lactamase. The 38 bp inverted repeats flanking Tn3 appear to be absolutely required in cis for Tn3 to transpose. Genetic data suggest that Tn3 contains a third site (Gill et al., 1978), designated IRS (internal resolution site), whose absence results in the insertion of two complete copies of Tn3 as direct repeats into the recipient DNA. We suggest that these direct repeats of complete copies of Tn3 are intermediates in transposition, and that the IRS site is required for recombination and subsequent segregation of the direct repeats to leave a single copy of Tn3 (Gill et al., 1978). A 23 nucleotide sequence within the amino terminus of the transposase which shares strong sequence homology with the inverted repeat may be the internal resolution site.

Original languageEnglish (US)
Pages (from-to)1153-1163
Number of pages11
JournalCell
Volume18
Issue number4
DOIs
StatePublished - 1979
Externally publishedYes

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Nucleic Acid Repetitive Sequences
DNA sequences
DNA Sequence Analysis
Transposases
Nucleotides
Genes
Amino Acids
Peptides
Mutation
Molecular Weight
Molecular weight
Sequence Homology
Genetic Recombination
Phenotype
DNA
Proteins

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

DNA sequence analysis of the transposon Tn3 : Three genes and three sites involved in transposition of Tn3. / Heffron, Fred; McCarthy, Brian J.; Ohtsubo, Hisako; Ohtsubo, Eiichi.

In: Cell, Vol. 18, No. 4, 1979, p. 1153-1163.

Research output: Contribution to journalArticle

Heffron, Fred ; McCarthy, Brian J. ; Ohtsubo, Hisako ; Ohtsubo, Eiichi. / DNA sequence analysis of the transposon Tn3 : Three genes and three sites involved in transposition of Tn3. In: Cell. 1979 ; Vol. 18, No. 4. pp. 1153-1163.
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