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PMID: 1647013 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

A covalent complex between retroviral integrase and nicked substrate DNA.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 88 ·No. 11 ·1991-06-01 ·Pages 4695-9

Katzman M, Mack JP, Skalka AM, Leis J

Abstract

Purified retroviral integrase (IN) from avian sarcoma-leukosis viruses can appropriately process the termini of linear viral DNA, cleave host DNA in a sequence-independent manner, and catalyze integrative recombination; an exogenous source of energy is not required for these reactions. Using DNA substrates containing radioactive phosphate groups, we demonstrate that IN becomes covalently joined to the new 5' phosphate ends of DNA produced at sites of cleavage. Most of the phosphodiester linkages between IN and DNA involve serine, but some involve threonine. Computer-assisted alignment of 80 retroviral and retrotransposon IN sequences identified one serine that is conserved in all of these proteins and three less-conserved threonine residues. These results identify candidate active-site residues and provide support for the participation of a covalent IN-DNA intermediate in retroviral integration.

MeSH Terms
Amino Acid Sequence Amino Acids/analysis Avian Myeloblastosis Virus/enzymology Avian Sarcoma Viruses/enzymology Base Sequence Binding Sites DNA Nucleotidyltransferases/metabolism DNA Transposable Elements DNA, Viral/metabolism Integrases Molecular Sequence Data Oligodeoxyribonucleotides/chemical synthesis,metabolism Phylogeny Protein Binding Retroviridae/enzymology,genetics Sequence Homology, Nucleic Acid Substrate Specificity
Chemicals
Amino Acids DNA Transposable Elements DNA, Viral Oligodeoxyribonucleotides DNA Nucleotidyltransferases Integrases
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Katzman M
Department of Medicine, Milton S. Hershey Medical Center, Pennsylvania State University College of Medicine, Hershey 17033.
Mack J P
Skalka A M
Leis J
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.88.11.4695
Published
1991-06-01
Pages
4695-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC51732
Subset
IM
Grants
NCI NIH HHS · CA 06927 · United States
NCI NIH HHS · CA 38046 · United States
NCI NIH HHS · CA 49042 · United States
External Links
PubMed source DOI: 10.1073/pnas.88.11.4695 PMC Full Text View journal details
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