Regulation of directionality in bacteriophage λ site-specific recombination

Structure of the Xis protein

My D. Sam, Christie V. Papagiannis, Kevin M. Connolly, Leah Corselli, Junji Iwahara, James Lee, Martin Phillips, Jonathan M. Wojciak, Reid C. Johnson, Robert T. Clubb

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Upon induction of a bacteriophage λ lysogen, a site-specific recombination reaction excises the phage genome from the chromosome of its bacterial host. A critical regulator of this process is the phage-encoded excisionase (Xis) protein, which functions both as a DNA architectural factor and by cooperatively recruiting integrase to an adjacent binding site specifically required for excision. Here we present the three-dimensional structure of Xis and the results of a structure-based mutagenesis study to define the molecular basis of its function. Xis adopts an unusual "winged"-helix motif that is modeled to interact with the major- and minor-grooves of its binding site through a single α-helix and loop structure ("wing"), respectively. The C-terminal tail of Xis, which is required for cooperative binding with integrase, is unstructured in the absence of DNA. We propose that asymmetric bending of DNA by Xis positions its unstructured C-terminal tail for direct contacts with the N-terminal DNA-binding domain of integrase and that an ensuing disordered to ordered transition of the tail may act to stabilize the formation of the tripartite integrase-Xis-DNA complex required for phage excision.

Original languageEnglish (US)
Pages (from-to)791-805
Number of pages15
JournalJournal of Molecular Biology
Volume324
Issue number4
DOIs
StatePublished - 2002
Externally publishedYes

Fingerprint

Integrases
Bacteriophages
Genetic Recombination
DNA
Proteins
Binding Sites
Bacterial Chromosomes
Mutagenesis
Genome

Keywords

  • NMR
  • Phage excision
  • Protein-DNA interactions
  • Site-specific DNA recombination
  • Structure

ASJC Scopus subject areas

  • Virology

Cite this

Regulation of directionality in bacteriophage λ site-specific recombination : Structure of the Xis protein. / Sam, My D.; Papagiannis, Christie V.; Connolly, Kevin M.; Corselli, Leah; Iwahara, Junji; Lee, James; Phillips, Martin; Wojciak, Jonathan M.; Johnson, Reid C.; Clubb, Robert T.

In: Journal of Molecular Biology, Vol. 324, No. 4, 2002, p. 791-805.

Research output: Contribution to journalArticle

Sam, MD, Papagiannis, CV, Connolly, KM, Corselli, L, Iwahara, J, Lee, J, Phillips, M, Wojciak, JM, Johnson, RC & Clubb, RT 2002, 'Regulation of directionality in bacteriophage λ site-specific recombination: Structure of the Xis protein', Journal of Molecular Biology, vol. 324, no. 4, pp. 791-805. https://doi.org/10.1016/S0022-2836(02)01150-6
Sam, My D. ; Papagiannis, Christie V. ; Connolly, Kevin M. ; Corselli, Leah ; Iwahara, Junji ; Lee, James ; Phillips, Martin ; Wojciak, Jonathan M. ; Johnson, Reid C. ; Clubb, Robert T. / Regulation of directionality in bacteriophage λ site-specific recombination : Structure of the Xis protein. In: Journal of Molecular Biology. 2002 ; Vol. 324, No. 4. pp. 791-805.
@article{3ec7c398f10e4e1786fcae2c285ae236,
title = "Regulation of directionality in bacteriophage λ site-specific recombination: Structure of the Xis protein",
abstract = "Upon induction of a bacteriophage λ lysogen, a site-specific recombination reaction excises the phage genome from the chromosome of its bacterial host. A critical regulator of this process is the phage-encoded excisionase (Xis) protein, which functions both as a DNA architectural factor and by cooperatively recruiting integrase to an adjacent binding site specifically required for excision. Here we present the three-dimensional structure of Xis and the results of a structure-based mutagenesis study to define the molecular basis of its function. Xis adopts an unusual {"}winged{"}-helix motif that is modeled to interact with the major- and minor-grooves of its binding site through a single α-helix and loop structure ({"}wing{"}), respectively. The C-terminal tail of Xis, which is required for cooperative binding with integrase, is unstructured in the absence of DNA. We propose that asymmetric bending of DNA by Xis positions its unstructured C-terminal tail for direct contacts with the N-terminal DNA-binding domain of integrase and that an ensuing disordered to ordered transition of the tail may act to stabilize the formation of the tripartite integrase-Xis-DNA complex required for phage excision.",
keywords = "NMR, Phage excision, Protein-DNA interactions, Site-specific DNA recombination, Structure",
author = "Sam, {My D.} and Papagiannis, {Christie V.} and Connolly, {Kevin M.} and Leah Corselli and Junji Iwahara and James Lee and Martin Phillips and Wojciak, {Jonathan M.} and Johnson, {Reid C.} and Clubb, {Robert T.}",
year = "2002",
doi = "10.1016/S0022-2836(02)01150-6",
language = "English (US)",
volume = "324",
pages = "791--805",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "4",

}

TY - JOUR

T1 - Regulation of directionality in bacteriophage λ site-specific recombination

T2 - Structure of the Xis protein

AU - Sam, My D.

AU - Papagiannis, Christie V.

AU - Connolly, Kevin M.

AU - Corselli, Leah

AU - Iwahara, Junji

AU - Lee, James

AU - Phillips, Martin

AU - Wojciak, Jonathan M.

AU - Johnson, Reid C.

AU - Clubb, Robert T.

PY - 2002

Y1 - 2002

N2 - Upon induction of a bacteriophage λ lysogen, a site-specific recombination reaction excises the phage genome from the chromosome of its bacterial host. A critical regulator of this process is the phage-encoded excisionase (Xis) protein, which functions both as a DNA architectural factor and by cooperatively recruiting integrase to an adjacent binding site specifically required for excision. Here we present the three-dimensional structure of Xis and the results of a structure-based mutagenesis study to define the molecular basis of its function. Xis adopts an unusual "winged"-helix motif that is modeled to interact with the major- and minor-grooves of its binding site through a single α-helix and loop structure ("wing"), respectively. The C-terminal tail of Xis, which is required for cooperative binding with integrase, is unstructured in the absence of DNA. We propose that asymmetric bending of DNA by Xis positions its unstructured C-terminal tail for direct contacts with the N-terminal DNA-binding domain of integrase and that an ensuing disordered to ordered transition of the tail may act to stabilize the formation of the tripartite integrase-Xis-DNA complex required for phage excision.

AB - Upon induction of a bacteriophage λ lysogen, a site-specific recombination reaction excises the phage genome from the chromosome of its bacterial host. A critical regulator of this process is the phage-encoded excisionase (Xis) protein, which functions both as a DNA architectural factor and by cooperatively recruiting integrase to an adjacent binding site specifically required for excision. Here we present the three-dimensional structure of Xis and the results of a structure-based mutagenesis study to define the molecular basis of its function. Xis adopts an unusual "winged"-helix motif that is modeled to interact with the major- and minor-grooves of its binding site through a single α-helix and loop structure ("wing"), respectively. The C-terminal tail of Xis, which is required for cooperative binding with integrase, is unstructured in the absence of DNA. We propose that asymmetric bending of DNA by Xis positions its unstructured C-terminal tail for direct contacts with the N-terminal DNA-binding domain of integrase and that an ensuing disordered to ordered transition of the tail may act to stabilize the formation of the tripartite integrase-Xis-DNA complex required for phage excision.

KW - NMR

KW - Phage excision

KW - Protein-DNA interactions

KW - Site-specific DNA recombination

KW - Structure

UR - http://www.scopus.com/inward/record.url?scp=0036928819&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036928819&partnerID=8YFLogxK

U2 - 10.1016/S0022-2836(02)01150-6

DO - 10.1016/S0022-2836(02)01150-6

M3 - Article

VL - 324

SP - 791

EP - 805

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 4

ER -