Abstract
We developed a purified system for reproducing the nicking reaction at the site 59 base pairs upstream of the TraY protein binding site, sbyA, in the oriT region of plasmid R100. Nicking at oriT occurred efficiently in the presence of the plasmid-encoded proteins, Tral and TraY, integration host factor (IHF), and Mg2+, but inefficiently in the presence of the Tral protein and Mg2+. The products were complex DNA molecules with a protein covalent-ly linked with the 5' end of the nick in the strand, which is supposed to be transferred during conjugation. The same complex DNA molecules were formed in the presence of the Tral protein alone, indicating that the protein attached at the 5' end of the nick is the Tral protein. Stimulation of the nicking reaction by the TraY protein and by IHF, whose binding site has been mapped between the nicking site and sbyA, indicates that DNA bending is important in the formation of the complex including the Tral and TraY proteins at oriT.
| Original language | English |
|---|---|
| Pages (from-to) | 838-844 |
| Number of pages | 7 |
| Journal | Journal of Biochemistry |
| Volume | 116 |
| Issue number | 4 |
| Publication status | Published - Oct 1994 |
| Externally published | Yes |
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Keywords
- Conjugation
- Endonuclease
- Origin of transfer
- Sex factor
ASJC Scopus subject areas
- Statistics, Probability and Uncertainty
- Applied Mathematics
- Physiology (medical)
- Radiology Nuclear Medicine and imaging
- Molecular Biology
- Biochemistry
Cite this
Site- and strand-specific nicking at oriT of plasmid R100 in a purified system : Enhancement of the nicking activity of tral (helicase I) with traY and IHF. / Inamoto, Susumu; Fukuda, Hirokazu; Abo, Tatsuhiko; Ohtsubo, Eiichi.
In: Journal of Biochemistry, Vol. 116, No. 4, 10.1994, p. 838-844.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Site- and strand-specific nicking at oriT of plasmid R100 in a purified system
T2 - Enhancement of the nicking activity of tral (helicase I) with traY and IHF
AU - Inamoto, Susumu
AU - Fukuda, Hirokazu
AU - Abo, Tatsuhiko
AU - Ohtsubo, Eiichi
PY - 1994/10
Y1 - 1994/10
N2 - We developed a purified system for reproducing the nicking reaction at the site 59 base pairs upstream of the TraY protein binding site, sbyA, in the oriT region of plasmid R100. Nicking at oriT occurred efficiently in the presence of the plasmid-encoded proteins, Tral and TraY, integration host factor (IHF), and Mg2+, but inefficiently in the presence of the Tral protein and Mg2+. The products were complex DNA molecules with a protein covalent-ly linked with the 5' end of the nick in the strand, which is supposed to be transferred during conjugation. The same complex DNA molecules were formed in the presence of the Tral protein alone, indicating that the protein attached at the 5' end of the nick is the Tral protein. Stimulation of the nicking reaction by the TraY protein and by IHF, whose binding site has been mapped between the nicking site and sbyA, indicates that DNA bending is important in the formation of the complex including the Tral and TraY proteins at oriT.
AB - We developed a purified system for reproducing the nicking reaction at the site 59 base pairs upstream of the TraY protein binding site, sbyA, in the oriT region of plasmid R100. Nicking at oriT occurred efficiently in the presence of the plasmid-encoded proteins, Tral and TraY, integration host factor (IHF), and Mg2+, but inefficiently in the presence of the Tral protein and Mg2+. The products were complex DNA molecules with a protein covalent-ly linked with the 5' end of the nick in the strand, which is supposed to be transferred during conjugation. The same complex DNA molecules were formed in the presence of the Tral protein alone, indicating that the protein attached at the 5' end of the nick is the Tral protein. Stimulation of the nicking reaction by the TraY protein and by IHF, whose binding site has been mapped between the nicking site and sbyA, indicates that DNA bending is important in the formation of the complex including the Tral and TraY proteins at oriT.
KW - Conjugation
KW - Endonuclease
KW - Origin of transfer
KW - Sex factor
UR - http://www.scopus.com/inward/record.url?scp=0028034221&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0028034221&partnerID=8YFLogxK
M3 - Article
C2 - 7883759
AN - SCOPUS:0028034221
VL - 116
SP - 838
EP - 844
JO - Journal of Biochemistry
JF - Journal of Biochemistry
SN - 0021-924X
IS - 4
ER -