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PMID: 3520564 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Purification and properties of the mini-F plasmid-encoded E protein needed for autonomous replication control of the plasmid.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 12 ·1986-06-00 ·Pages 4109-13

Tokino T, Murotsu T, Matsubara K

Abstract

Mini-F plasmid encodes a protein, E protein, that is indispensable for its autonomous replication. We have constructed a plasmid that overproduces the E protein and have purified the protein to apparent homogeneity. Using nitrocellulose filter binding and nuclease digestion assays, we demonstrated that the E protein binds to three unique regions of the mini-F DNA sequence: the replication origin (ori2) and an incompatibility locus (incB), another incompatibility locus (incC), and the promoter for the E gene. These binding sites have a common 8-base-pair sequence. These findings suggest the direct role of the E protein in initiation of mini-F replication and copy number control. They are also in line with the in vivo evidence that the incompatibility phenotype caused by incB and incC DNA is due to titration of a factor(s) indispensable for replication and that the production of the E initiator protein of the mini-F plasmid is under autoregulatory control.

MeSH Terms

Amino Acid Sequence Bacterial Proteins/isolation & purification,metabolism Base Sequence DNA Replication DNA-Binding Proteins/isolation & purification,metabolism Deoxyribonuclease I Escherichia coli/genetics Exodeoxyribonucleases F Factor Repetitive Sequences, Nucleic Acid

Chemicals

Bacterial Proteins DNA-Binding Proteins Exodeoxyribonucleases exodeoxyribonuclease III Deoxyribonuclease I

Authors & Affiliations

3 authors, click to expand affiliations / ORCID

Tokino T

Murotsu T

Matsubara K

References (25)

25 references, click to expand

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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.83.12.4109

Published

1986-06-00

Pages

4109-13

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC323680

Subset

External Links

PubMed source DOI: 10.1073/pnas.83.12.4109 PMC Full Text View journal details

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Purification and properties of the mini-F plasmid-encoded E protein needed for autonomous replication control of the plasmid.

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