Cooperative binding of the E2 protein of bovine papillomavirus to adjacent E2-responsive sequences

Paolo Monini, Steven R. Grossman, Blake Pepinsky, Elliot Androphy, Laimonis A. Laimins

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

The DNA-binding properties of purified full-length E2 protein from bovine papillomavirus type 1 have been investigated by utilizing a quantitative gel shift analysis. By using a recombinant baculovirus which expresses the E2 open reading frame from the polyhedrin promoter, the full-length E2 protein was synthesized in insect cells and purified to homogeneity by using an E2 binding site (ACCGN4CGGT)-specific oligonucleotide column. The Kd of E2 binding to a 41-bp oligonucleotide containing a single binding site was found to be 2 × 10d M. When two binding sites were included on an oligonucleotide, cooperative binding to these sites by the E2 protein was observed. A cooperativity parameter of 8.5 was determined for E2 binding to two sites. An 86-amino-acid peptide encompassing the C terminus of the protein retains the ability to bind E2 binding sites with a Kd of 4 × 10-10 M but exhibits slight cooperativity of binding to two adjacent sites. A major determinant for cooperative binding of the full-length E2 protein is thus encoded by the N-terminal amino acids outside the minimal DNA binding domain.

Original languageEnglish (US)
Pages (from-to)2124-2130
Number of pages7
JournalJournal of Virology
Volume65
Issue number4
StatePublished - 1991
Externally publishedYes

Fingerprint

Bovine papillomavirus
cooperatives
Carrier Proteins
Binding Sites
binding sites
oligonucleotides
Oligonucleotides
Proteins
proteins
Bovine papillomavirus 1
Amino Acids
amino acids
DNA-binding domains
binding properties
Baculoviridae
DNA
Electrophoretic Mobility Shift Assay
Open Reading Frames
Insects
open reading frames

ASJC Scopus subject areas

  • Immunology

Cite this

Monini, P., Grossman, S. R., Pepinsky, B., Androphy, E., & Laimins, L. A. (1991). Cooperative binding of the E2 protein of bovine papillomavirus to adjacent E2-responsive sequences. Journal of Virology, 65(4), 2124-2130.

Cooperative binding of the E2 protein of bovine papillomavirus to adjacent E2-responsive sequences. / Monini, Paolo; Grossman, Steven R.; Pepinsky, Blake; Androphy, Elliot; Laimins, Laimonis A.

In: Journal of Virology, Vol. 65, No. 4, 1991, p. 2124-2130.

Research output: Contribution to journalArticle

Monini, P, Grossman, SR, Pepinsky, B, Androphy, E & Laimins, LA 1991, 'Cooperative binding of the E2 protein of bovine papillomavirus to adjacent E2-responsive sequences', Journal of Virology, vol. 65, no. 4, pp. 2124-2130.
Monini, Paolo ; Grossman, Steven R. ; Pepinsky, Blake ; Androphy, Elliot ; Laimins, Laimonis A. / Cooperative binding of the E2 protein of bovine papillomavirus to adjacent E2-responsive sequences. In: Journal of Virology. 1991 ; Vol. 65, No. 4. pp. 2124-2130.
@article{4e8c4b8602f6445d8500b179f1b76ddb,
title = "Cooperative binding of the E2 protein of bovine papillomavirus to adjacent E2-responsive sequences",
abstract = "The DNA-binding properties of purified full-length E2 protein from bovine papillomavirus type 1 have been investigated by utilizing a quantitative gel shift analysis. By using a recombinant baculovirus which expresses the E2 open reading frame from the polyhedrin promoter, the full-length E2 protein was synthesized in insect cells and purified to homogeneity by using an E2 binding site (ACCGN4CGGT)-specific oligonucleotide column. The Kd of E2 binding to a 41-bp oligonucleotide containing a single binding site was found to be 2 × 10d M. When two binding sites were included on an oligonucleotide, cooperative binding to these sites by the E2 protein was observed. A cooperativity parameter of 8.5 was determined for E2 binding to two sites. An 86-amino-acid peptide encompassing the C terminus of the protein retains the ability to bind E2 binding sites with a Kd of 4 × 10-10 M but exhibits slight cooperativity of binding to two adjacent sites. A major determinant for cooperative binding of the full-length E2 protein is thus encoded by the N-terminal amino acids outside the minimal DNA binding domain.",
author = "Paolo Monini and Grossman, {Steven R.} and Blake Pepinsky and Elliot Androphy and Laimins, {Laimonis A.}",
year = "1991",
language = "English (US)",
volume = "65",
pages = "2124--2130",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "4",

}

TY - JOUR

T1 - Cooperative binding of the E2 protein of bovine papillomavirus to adjacent E2-responsive sequences

AU - Monini, Paolo

AU - Grossman, Steven R.

AU - Pepinsky, Blake

AU - Androphy, Elliot

AU - Laimins, Laimonis A.

PY - 1991

Y1 - 1991

N2 - The DNA-binding properties of purified full-length E2 protein from bovine papillomavirus type 1 have been investigated by utilizing a quantitative gel shift analysis. By using a recombinant baculovirus which expresses the E2 open reading frame from the polyhedrin promoter, the full-length E2 protein was synthesized in insect cells and purified to homogeneity by using an E2 binding site (ACCGN4CGGT)-specific oligonucleotide column. The Kd of E2 binding to a 41-bp oligonucleotide containing a single binding site was found to be 2 × 10d M. When two binding sites were included on an oligonucleotide, cooperative binding to these sites by the E2 protein was observed. A cooperativity parameter of 8.5 was determined for E2 binding to two sites. An 86-amino-acid peptide encompassing the C terminus of the protein retains the ability to bind E2 binding sites with a Kd of 4 × 10-10 M but exhibits slight cooperativity of binding to two adjacent sites. A major determinant for cooperative binding of the full-length E2 protein is thus encoded by the N-terminal amino acids outside the minimal DNA binding domain.

AB - The DNA-binding properties of purified full-length E2 protein from bovine papillomavirus type 1 have been investigated by utilizing a quantitative gel shift analysis. By using a recombinant baculovirus which expresses the E2 open reading frame from the polyhedrin promoter, the full-length E2 protein was synthesized in insect cells and purified to homogeneity by using an E2 binding site (ACCGN4CGGT)-specific oligonucleotide column. The Kd of E2 binding to a 41-bp oligonucleotide containing a single binding site was found to be 2 × 10d M. When two binding sites were included on an oligonucleotide, cooperative binding to these sites by the E2 protein was observed. A cooperativity parameter of 8.5 was determined for E2 binding to two sites. An 86-amino-acid peptide encompassing the C terminus of the protein retains the ability to bind E2 binding sites with a Kd of 4 × 10-10 M but exhibits slight cooperativity of binding to two adjacent sites. A major determinant for cooperative binding of the full-length E2 protein is thus encoded by the N-terminal amino acids outside the minimal DNA binding domain.

UR - http://www.scopus.com/inward/record.url?scp=0026078079&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026078079&partnerID=8YFLogxK

M3 - Article

C2 - 1848322

AN - SCOPUS:0026078079

VL - 65

SP - 2124

EP - 2130

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 4

ER -