期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 107, 期 22, 页码 10014-10019出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0912615107
关键词
MuA transposase; Mu DNA transposition; ClpX; retroviral integration; post-integration repair
资金
- National Institutes of Health [GM 33247]
- Robert Welch Foundation [F-1351]
Phage Mu transposes by two distinct pathways depending on the specific stage of its life cycle. A common. strand transfer intermediate is resolved differentially in the two pathways. During lytic growth, the. intermediate is resolved by replication of Mu initiated within the flanking target DNA; during integration of infecting Mu, it is resolved without replication, by removal and repair of DNA from a previous host that is still attached to the ends of the incoming Mu genome. We have discovered that the cryptic endonuclease activity reported for the isolated C-terminal domain of the transposase MuA [Wu Z, Chaconas G (1995) A novel DNA binding and nuclease activity in domain III of Mu transposase: Evidence for a catalytic region involved in donor cleavage. EMBO J 14:3835-3843], which is not observed in the full-length protein or in the assembled transpososome in vitro, is required in vivo for removal of the attached host DNA or 5' flap after the infecting Mu genome has integrated into the E. coli chromosome. Efficient flap removal also requires the host protein ClpX, which is known to interact with the C-terminus of MuA to remodel the transpososome for replication. We hypothesize that ClpX constitutes part of a highly regulated mechanism that unmasks the cryptic nuclease activity of MuA specifically in the repair pathway.
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