Home LiteratureArticle Details
PMID: 3461445 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Sequence-specific interaction between the replication initiator protein of plasmid pT181 and its origin of replication.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 15 ·1986-08-00 ·Pages 5484-8

Koepsel RR, Murray RW, Khan SA

Abstract

The replication of the pT181 plasmid is dependent on the plasmid-encoded initiator protein RepC. We have previously shown that RepC protein has sequence-specific endonuclease and topoisomerase-like activities. In this paper we demonstrate that this initiator protein has sequence-specific DNA-binding properties. Based on filter binding of plasmid restriction fragments, RepC protein specifically recognizes only the pT181 origin region. Using DNase I and neocarzinostatin "footprinting" techniques, we show that RepC protein specifically binds to a 32-base-pair sequence within the origin that is part of the initiator cistron. Using dimethyl sulfate as a chemical probe, we have identified the purine residues that interact with the initiator protein. The features of the DNA region that interacts with RepC protein include sequences with the potential to form Z DNA and/or hairpin structures. The specific DNA-protein interaction at the origin may be critical in the initiation of pT181 DNA replication by RepC protein in association with other host initiation proteins.

MeSH Terms
Bacterial Proteins/genetics Base Sequence Binding Sites DNA Replication DNA, Bacterial/biosynthesis DNA-Binding Proteins/genetics Hydrolysis Plasmids Staphylococcus aureus/genetics Sulfuric Acid Esters
Chemicals
Bacterial Proteins DNA, Bacterial DNA-Binding Proteins RepC protein, Staphylococcus aureus Sulfuric Acid Esters dimethyl sulfate
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Koepsel R R
Murray R W
Khan S A
References (33)
33 references, click to expand
  1. Purification of pT181-encoded repC protein required for the initiation of plasmid replication.
    J Biol Chem. 1985 Jul 15;260(14):8571-7 PMID: 2989292
  2. Interaction of the plasmid R6K-encoded replication initiator protein with its binding sites on DNA.
    Cell. 1983 Aug;34(1):125-34 PMID: 6224568
  3. The replication initiator protein of plasmid R6K tagged with beta-galactosidase shows sequence-specific DNA-binding.
    Cell. 1983 Jan;32(1):131-40 PMID: 6297781
  4. Mutations in direct repeat sequences and in a conserved sequence adjacent to the repeats result in a defective replication origin in plasmid R6K.
    Proc Natl Acad Sci U S A. 1985 Mar;82(5):1480-4 PMID: 3883361
  5. Nine unique repeating sequences in a region essential for replication and incompatibility of the mini-F plasmid.
    Gene. 1981 Nov;15(2-3):257-71 PMID: 7028572
  6. Coding sequence for the pT181 repC product: a plasmid-coded protein uniquely required for replication.
    Proc Natl Acad Sci U S A. 1982 Jul;79(13):4108-12 PMID: 6287465
  7. Nucleotide sequence of the region of an origin of replication of the antibiotic resistance plasmid R6K.
    Proc Natl Acad Sci U S A. 1979 Mar;76(3):1150-4 PMID: 375227
  8. Autoregulation of simian virus 40 gene A by T antigen.
    Proc Natl Acad Sci U S A. 1976 Sep;73(9):3083-7 PMID: 184459
  9. Purified bacteriophage lambda O protein binds to four repeating sequences at the lambda replication origin.
    Nucleic Acids Res. 1981 Apr 24;9(8):1789-99 PMID: 6264392
  10. The replication initiator protein of plasmid pT181 has sequence-specific endonuclease and topoisomerase-like activities.
    Proc Natl Acad Sci U S A. 1985 Oct;82(20):6845-9 PMID: 2995991
  11. The replication initiator protein of plasmid pSC101 is a transcriptional repressor of its own cistron.
    Proc Natl Acad Sci U S A. 1985 Apr;82(8):2252-6 PMID: 2986108
  12. Plasmid pT181 replication is regulated by two countertranscripts.
    Proc Natl Acad Sci U S A. 1985 Feb;82(3):638-42 PMID: 2579377
  13. The binding site on SV40 DNA for a T antigen-related protein.
    Cell. 1978 Jan;13(1):165-79 PMID: 202398
  14. Control of pT181 replication I. The pT181 copy control function acts by inhibiting the synthesis of a replication protein.
    EMBO J. 1984 Oct;3(10):2399-405 PMID: 6499834
  15. Negatively supercoiled plasmids contain left-handed Z-DNA segments as detected by specific antibody binding.
    Cell. 1982 Dec;31(2 Pt 1):309-18 PMID: 7159926
  16. Nucleotide sequence of the region of the origin of replication of the broad host range plasmid RK2.
    Mol Gen Genet. 1981;181(1):8-12 PMID: 6261086
  17. DNA-protein interaction at the origin of DNA replication of the plasmid pSC101.
    Cell. 1983 Dec;35(2 Pt 1):495-502 PMID: 6317193
  18. P1 plasmid replication: multiple functions of RepA protein at the origin.
    Proc Natl Acad Sci U S A. 1985 May;82(9):2588-92 PMID: 3857601
  19. Right-handed and left-handed DNA: conformational information in genetic material.
    Cold Spring Harb Symp Quant Biol. 1983;47 Pt 1:1-12 PMID: 6574832
  20. An amino-terminal fragment of lac repressor binds specifically to lac operator.
    Proc Natl Acad Sci U S A. 1978 Dec;75(12):5851-4 PMID: 16592594
  21. The dnaA protein complex with the E. coli chromosomal replication origin (oriC) and other DNA sites.
    Cell. 1984 Oct;38(3):889-900 PMID: 6091903
  22. Control of pT181 replication II. Mutational analysis.
    EMBO J. 1984 Oct;3(10):2407-14 PMID: 6437809
  23. Replication initiator protein of plasmid R6K autoregulates its own synthesis at the transcriptional step.
    Proc Natl Acad Sci U S A. 1985 May;82(9):2574-8 PMID: 3857600
  24. Complete nucleotide sequence of pT181, a tetracycline-resistance plasmid from Staphylococcus aureus.
    Plasmid. 1983 Nov;10(3):251-9 PMID: 6657777
  25. DNAse footprinting: a simple method for the detection of protein-DNA binding specificity.
    Nucleic Acids Res. 1978 Sep;5(9):3157-70 PMID: 212715
  26. Identification of the minimal essential region for the replication origin of miniF plasmid.
    Mol Gen Genet. 1984;196(2):373-8 PMID: 6593565
  27. Plasmid R6K DNA replication. III. Regulatory properties of the pi initiation protein.
    J Mol Biol. 1982 Oct 15;161(1):57-76 PMID: 6818353
  28. Replication of plasmid pT181 DNA in vitro: requirement for a plasmid-encoded product.
    Proc Natl Acad Sci U S A. 1981 Aug;78(8):4902-6 PMID: 6946436
  29. Regulation of plasmid replication.
    Microbiol Rev. 1984 Mar;48(1):1-23 PMID: 6201704
  30. Autoregulation of the DNA replication gene dnaA in E. coli K-12.
    Cell. 1985 Jan;40(1):159-69 PMID: 2981626
  31. Sequencing end-labeled DNA with base-specific chemical cleavages.
    Methods Enzymol. 1980;65(1):499-560 PMID: 6246368
  32. Replication initiated at the origin (oriC) of the E. coli chromosome reconstituted with purified enzymes.
    Cell. 1984 Aug;38(1):183-90 PMID: 6088063
  33. Functional origin of replication of pT181 plasmid DNA is contained within a 168-base-pair segment.
    Proc Natl Acad Sci U S A. 1982 Aug;79(15):4580-4 PMID: 6956881
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.83.15.5484
Published
1986-08-00
Pages
5484-8
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC386311
Subset
IM
Grants
NIGMS NIH HHS · GM 31685 · United States
External Links
PubMed source DOI: 10.1073/pnas.83.15.5484 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2