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PMID: 8428593 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Origin recognition specificity in pT181 plasmids is determined by a functionally asymmetric palindromic DNA element.

The EMBO journal ·Vol. 12 ·No. 1 ·1993-01-00 ·Pages 45-52

Wang PZ, Projan SJ, Henriquez V, Novick RP

Abstract

The leading strand replication origin of pT181 plasmids consists of two adjacent inverted repeat elements (IR-II and IR-III), which are involved in origin recognition by the initiator (Rep) protein. The conserved core element, IR-II, which contains the initiation nick site, is induced by Rep to form a cruciform structure, probably the primary substrate for the initiation of rolling circle replication. The divergent repeat, IR-III, constitutes the determinant of origin recognition specificity. We show here that the distal arm of IR-III is not required for sequence-specific recognition, whereas the proximal arm and central region are required. Since the initiator is dimeric, we presume that it binds symmetrically to IR-III. A unique type of DNA-protein interaction is proposed, in which the lack of sequence requirement for the distal arm is a consequence of binding to the adjacent IR-II, which thereby polarizes the stringency of binding to the two arms of IR-III. In addition, genetic evidence indicates that both the spacing and the phasing of IR-II to IR-III are crucial for function and that the central segment of IR-III may serve to position the two flanking half-sites for optimal interaction of Rep with IR-III.

Related Genes
oriC repC
MeSH Terms
Amino Acid Sequence Bacterial Proteins/genetics,metabolism Base Sequence Cloning, Molecular DNA Replication DNA, Bacterial/genetics,metabolism DNA-Binding Proteins/genetics,metabolism Escherichia coli/genetics,metabolism Genes, Bacterial Models, Structural Molecular Sequence Data Mutagenesis, Site-Directed Nucleic Acid Conformation Plasmids Repetitive Sequences, Nucleic Acid Replicon Restriction Mapping Sequence Homology, Amino Acid
Chemicals
Bacterial Proteins DNA, Bacterial DNA-Binding Proteins RepC protein, Staphylococcus aureus
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Wang P Z
Department of Plasmid Biology and Applied Genetics, Public Health Research Institute of the City of New York, Inc., NY 10016.
Projan S J
Henriquez V
Novick R P
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1993-01-00
Pages
45-52
Language
English
Region
England
NLM ID
8208664
PMCID
PMC413174
Subset
IM
Grants
NIGMS NIH HHS · GM14372 · United States
Databases
GENBANK
L09677, L09678, L09679, L09680, L09681, L09682, L09683, X02529, X06627, X07371
External Links
PubMed source PMC Full Text View journal details
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