Bacteriophage P22 Mnt repressor. DNA binding and effects on transcription in vitro

Andrew K. Vershon; Sha Mei Liao; William R. McClure; Robert T. Sauer

doi:10.1016/0022-2836(87)90652-8

Bacteriophage P22 Mnt repressor. DNA binding and effects on transcription in vitro

Andrew K. Vershon, Sha Mei Liao, William R. McClure, Robert T. Sauer

Research output: Contribution to journal › Article › peer-review

57 Scopus citations

Abstract

We have examined the binding of Mnt repressor to operator DNA in vitro and have determined how this binding affects the level of transcription from two nearby promoters, P_ant and P_mnt. Mnt binds to a region of DNA that overlaps the startpoint of transcription of P_ant and the -35 region of P_mnt. Mnt represses transcription in vitro from P_ant and enhances transcription from P_mnt. Protection and interference experiments show that Mnt binds to a single, 17 base-pair operator site. The operator sequence and the protein-DNA contacts are symmetric. Mnt makes major groove contacts on both faces of the operator DNA. At pH 7.5, 200 mm-KCl, 22 °C, the Mnt tetramer binds operator with high affinity Kd = 2.2 × 10^-11m and the protein-DNA complex is quite stable (t_{1 2} = 48 min). Operator binding shows large dependencies on pH, salt concentration, and temperature.

Original language	English (US)
Pages (from-to)	311-322
Number of pages	12
Journal	Journal of molecular biology
Volume	195
Issue number	2
DOIs	https://doi.org/10.1016/0022-2836(87)90652-8
State	Published - May 20 1987
Externally published	Yes

All Science Journal Classification (ASJC) codes

Structural Biology
Molecular Biology

Access to Document

10.1016/0022-2836(87)90652-8

Cite this

@article{d6e5d2e800be495a8635b1ff93f9209a,

title = "Bacteriophage P22 Mnt repressor. DNA binding and effects on transcription in vitro",

abstract = "We have examined the binding of Mnt repressor to operator DNA in vitro and have determined how this binding affects the level of transcription from two nearby promoters, Pant and Pmnt. Mnt binds to a region of DNA that overlaps the startpoint of transcription of Pant and the -35 region of Pmnt. Mnt represses transcription in vitro from Pant and enhances transcription from Pmnt. Protection and interference experiments show that Mnt binds to a single, 17 base-pair operator site. The operator sequence and the protein-DNA contacts are symmetric. Mnt makes major groove contacts on both faces of the operator DNA. At pH 7.5, 200 mm-KCl, 22 °C, the Mnt tetramer binds operator with high affinity Kd = 2.2 × 10-11m and the protein-DNA complex is quite stable (t 1 2 = 48 min). Operator binding shows large dependencies on pH, salt concentration, and temperature.",

author = "Vershon, {Andrew K.} and Liao, {Sha Mei} and McClure, {William R.} and Sauer, {Robert T.}",

note = "Funding Information: We thank Mimi Susskind and Phil Youderian for strains and helpful discussion, and Gary Quigley for help with computer graphics. This work was supported by NIH grants AI-16892 (to R.T.S.) and GM-30375 (to W.R.M.).",

year = "1987",

month = may,

day = "20",

doi = "10.1016/0022-2836(87)90652-8",

language = "English (US)",

volume = "195",

pages = "311--322",

journal = "Journal of molecular biology",

issn = "0022-2836",

publisher = "Academic Press Inc.",

number = "2",

}

TY - JOUR

T1 - Bacteriophage P22 Mnt repressor. DNA binding and effects on transcription in vitro

AU - Vershon, Andrew K.

AU - Liao, Sha Mei

AU - McClure, William R.

AU - Sauer, Robert T.

N1 - Funding Information: We thank Mimi Susskind and Phil Youderian for strains and helpful discussion, and Gary Quigley for help with computer graphics. This work was supported by NIH grants AI-16892 (to R.T.S.) and GM-30375 (to W.R.M.).

PY - 1987/5/20

Y1 - 1987/5/20

N2 - We have examined the binding of Mnt repressor to operator DNA in vitro and have determined how this binding affects the level of transcription from two nearby promoters, Pant and Pmnt. Mnt binds to a region of DNA that overlaps the startpoint of transcription of Pant and the -35 region of Pmnt. Mnt represses transcription in vitro from Pant and enhances transcription from Pmnt. Protection and interference experiments show that Mnt binds to a single, 17 base-pair operator site. The operator sequence and the protein-DNA contacts are symmetric. Mnt makes major groove contacts on both faces of the operator DNA. At pH 7.5, 200 mm-KCl, 22 °C, the Mnt tetramer binds operator with high affinity Kd = 2.2 × 10-11m and the protein-DNA complex is quite stable (t 1 2 = 48 min). Operator binding shows large dependencies on pH, salt concentration, and temperature.

AB - We have examined the binding of Mnt repressor to operator DNA in vitro and have determined how this binding affects the level of transcription from two nearby promoters, Pant and Pmnt. Mnt binds to a region of DNA that overlaps the startpoint of transcription of Pant and the -35 region of Pmnt. Mnt represses transcription in vitro from Pant and enhances transcription from Pmnt. Protection and interference experiments show that Mnt binds to a single, 17 base-pair operator site. The operator sequence and the protein-DNA contacts are symmetric. Mnt makes major groove contacts on both faces of the operator DNA. At pH 7.5, 200 mm-KCl, 22 °C, the Mnt tetramer binds operator with high affinity Kd = 2.2 × 10-11m and the protein-DNA complex is quite stable (t 1 2 = 48 min). Operator binding shows large dependencies on pH, salt concentration, and temperature.

UR - http://www.scopus.com/inward/record.url?scp=0023660373&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023660373&partnerID=8YFLogxK

U2 - 10.1016/0022-2836(87)90652-8

DO - 10.1016/0022-2836(87)90652-8

M3 - Article

C2 - 3656414

AN - SCOPUS:0023660373

SN - 0022-2836

VL - 195

SP - 311

EP - 322

JO - Journal of molecular biology

JF - Journal of molecular biology

IS - 2

ER -

Bacteriophage P22 Mnt repressor. DNA binding and effects on transcription in vitro

Abstract

All Science Journal Classification (ASJC) codes

Access to Document

Other files and links

Fingerprint

Cite this