Characterization of a conjugative transposon integrase, IntDOT.

Characterization of a conjugative transposon integrase, IntDOT. Research Abstract Details 

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Characterization of a conjugative transposon integrase, IntDOT. Abstract Text:

Karolina Malanowska,Abigail A Salyers,Jeffrey F Gardner,

Sequence analysis revealed that the integrase of the Bacteroides conjugative transposon CTnDOT (IntDOT) might be a member of the tyrosine recombinase family because IntDOT has five of six highly conserved residues found in the catalytic domains of tyrosine recombinases. Yet, IntDOT catalyses a reaction that appears to differ in some respects from well-studied tyrosine recombinases such as that of phage lambda. To assess the importance of the conserved residues, we changed residues in IntDOT that align with conserved residues in tyrosine recombinases. Some substitutions resulted in a complete loss or significant decrease of integration activity in vivo. The ability of the mutant proteins to cleave and ligate CTnDOT attachment site (attDOT) DNA in vitro in general paralleled the in vivo results, but the H345A mutant, which had a wild-type level of integration in vivo, exhibited a slightly lower level of cleavage and ligation in vitro. Our results confirm the hypothesis that IntDOT belongs to the tyrosine recombinase family, but the catalytic core of the protein seems to have somewhat different organization. Previous DNA sequence analyses showed that CTnDOT att sites contain 5 bp non-homologous coupling sequences which were assumed to define the putative staggered sites of cleavage. However, cleavage assays showed that one of the cleavage sites is 2 bp away from the junction of CTnDOT and coupling sequence DNA. The site is in a region of homology that is conserved in CTnDOT att sites.

Characterization of a conjugative transposon integrase, IntDOT. Publishing Authors By Initials

K Malanowska,AA Salyers,JF Gardner,

For similar amino acids, peptides, and proteins: amino acids: amino acids, cyclic: amino acids, aromatic: tyrosine research abstracts see: amino acids, peptides, and proteins: amino acids: amino acids, cyclic: amino acids, aromatic: tyrosine research

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Characterization of a conjugative transposon integrase, IntDOT. Journal Published:

PUBLICATION TYPE: Research Support, N.I.H., Extr

Journal: Molecular microbiology

VOLUME: 60

Page Numbers: 1228-40

Journal Abbreviation: Mol. Microbiol.

ISSN: 0950-382X

DAY: 3

MONTH: Jun

YEAR: 2006

Characterization of a conjugative transposon integrase, IntDOT. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 8712028

Characterization of a conjugative transposon integrase, IntDOT. Keywords Mesh Terms:

KEYWORDS: Tyrosine

MESH TERMS: metabolism

Chemical & Substance for Abstract: Characterization of a conjugative transposon integrase, IntDOT. Information

Substance Name: Integrases

Registry Number: EC 2.7.7.-

Grant and Affiliation Information for Characterization of a conjugative transposon integrase, IntDOT.

AFFILIATION: Department of Microbiology, University of Illinois, Urbana, IL 61801, USA. malanows@uiuc.edu

Country: England

AGENCY: United States NIGMS

GRANT: GM 28717

ACRONYM: GM

MEDLINETA: Mol Microbiol

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