Abstract:
:The replication origin of the broad-host-range plasmid RK2, oriV, contains four DnaA boxes, which bind the DnaA protein isolated from Escherichia coli. Using a transformation assay, mutational analysis of these boxes showed a differential requirement for replication in different Gram-negative bacteria. DnaA boxes 3 and 4 were required in E. coli and Pseudomonas putidabut not as strictly in Azotobacter vinelandii and not at all in P. aeruginosa. In vitro replication results using an extract prepared from E. coli demonstrated that the activity of origin derivatives containing mutations in boxes 3 or 4 or a deletion of all four DnaA boxes could be restored by the addition of increasing amounts of purified DnaA protein. High levels of DnaA protein in the presence of the TrfA protein also resulted in the stimulation of open complex formation and DnaB helicase loading on oriV, even in the absence of the four DnaA boxes. These observations at least raise the possibility that an alternative mechanism of initiation of oriV is being used in the absence of the four DnaA boxes and that this mechanism may be similar to that used in P. aeruginosa, which does not require these four DnaA boxes for replication.
journal_name
Mol Microbioljournal_title
Molecular microbiologyauthors
Doran KS,Helinski DR,Konieczny Idoi
10.1046/j.1365-2958.1999.01491.xsubject
Has Abstractpub_date
1999-08-01 00:00:00pages
490-8issue
3eissn
0950-382Xissn
1365-2958pii
mole1491journal_volume
33pub_type
杂志文章abstract::We report a novel strategy for selecting mutations that mislocalize lipoproteins within the Escherichia coli cell envelope and describe the mutants obtained. A strain carrying a deletion of the chromosomal malE gene, coding for the periplasmic maltose-binding protein (MalE), cannot use maltose unless a wild-type copy ...
journal_title:Molecular microbiology
pub_type: 杂志文章
doi:10.1046/j.1365-2958.1999.01647.x
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journal_title:Molecular microbiology
pub_type: 杂志文章
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更新日期:1995-08-01 00:00:00
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pub_type: 杂志文章
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pub_type: 评论,杂志文章
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更新日期:2008-11-01 00:00:00
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pub_type: 杂志文章
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更新日期:2007-07-01 00:00:00
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pub_type: 杂志文章
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pub_type: 杂志文章,评审
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更新日期:2020-09-29 00:00:00