Binding-induced Stabilization and Assembly of the Phage P22 Tail Accessory Factor Gp4

Adam S. Olia, Jawdat Al-Bassam, Danella A. Winn-Stapley, Lisa Joss, Sherwood R. Casjens, Gino Cingolani

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

To infect and replicate, bacteriophage P22 injects its 43 kbp genome across the cell wall of Salmonella enterica serovar Typhimurium. The attachment of phage P22 to the host cell as well as the injection of the viral DNA into the host is mediated by the virion's tail complex. This 2.8 MDa molecular machine is formed by five proteins, which include the portal protein gp1, the adhesion tailspike protein gp9, and three tail accessory factors: gp4, gp10, gp26. We have isolated the tail accessory factor gp4 and characterized its structure and binding interactions with portal protein. Interestingly, gp4 exists in solution as a monomer, which displays an exceedingly low structural stability (Tm 34 °C). Unfolded gp4 is prone to aggregation within a narrow range of temperatures both in vitro and in Salmonella extracts. In the virion the thermal unfolding of gp4 is prevented by the interaction with the dodecameric portal protein, which stabilizes the structure of gp4 and suppresses unfolded gp4 from irreversibly aggregating in the Salmonella milieu. The structural stabilization of gp4 is accompanied by the concomitant oligomerization of the protein to form a ring of 12 subunits bound to the lower end of the portal ring. The interaction of gp4 with portal protein is complex and likely involves the distinct binding of two non-equivalent sets of six gp4 proteins. Binding of the first set of six gp4 equivalents to dodecameric portal protein yields a gp(1)12:gp(4)6 assembly intermediate, which is stably populated at 30 °C and can be resolved by native gel electrophoresis. The final product of the assembly reaction is a bi-dodecameric gp(1)12:gp(4)12 complex, which appears hollow by electron microscopy, suggesting that gp4 does not physically plug the DNA entry/exit channel, but acts as a structural adaptor for the other tail accessory factors: gp10 and gp26.

Original languageEnglish (US)
Pages (from-to)558-576
Number of pages19
JournalJournal of Molecular Biology
Volume363
Issue number2
DOIs
StatePublished - Oct 20 2006
Externally publishedYes

Fingerprint

Bacteriophage P22
Proteins
Salmonella
Virion
Salmonella enterica
Viral DNA
Cell Wall
Electrophoresis
Electron Microscopy
Hot Temperature
Gels
Genome

Keywords

  • bacteriophage P22
  • gp10
  • gp26
  • gp4
  • portal protein

ASJC Scopus subject areas

  • Virology

Cite this

Olia, A. S., Al-Bassam, J., Winn-Stapley, D. A., Joss, L., Casjens, S. R., & Cingolani, G. (2006). Binding-induced Stabilization and Assembly of the Phage P22 Tail Accessory Factor Gp4. Journal of Molecular Biology, 363(2), 558-576. https://doi.org/10.1016/j.jmb.2006.08.014

Binding-induced Stabilization and Assembly of the Phage P22 Tail Accessory Factor Gp4. / Olia, Adam S.; Al-Bassam, Jawdat; Winn-Stapley, Danella A.; Joss, Lisa; Casjens, Sherwood R.; Cingolani, Gino.

In: Journal of Molecular Biology, Vol. 363, No. 2, 20.10.2006, p. 558-576.

Research output: Contribution to journalArticle

Olia, AS, Al-Bassam, J, Winn-Stapley, DA, Joss, L, Casjens, SR & Cingolani, G 2006, 'Binding-induced Stabilization and Assembly of the Phage P22 Tail Accessory Factor Gp4', Journal of Molecular Biology, vol. 363, no. 2, pp. 558-576. https://doi.org/10.1016/j.jmb.2006.08.014
Olia AS, Al-Bassam J, Winn-Stapley DA, Joss L, Casjens SR, Cingolani G. Binding-induced Stabilization and Assembly of the Phage P22 Tail Accessory Factor Gp4. Journal of Molecular Biology. 2006 Oct 20;363(2):558-576. https://doi.org/10.1016/j.jmb.2006.08.014
Olia, Adam S. ; Al-Bassam, Jawdat ; Winn-Stapley, Danella A. ; Joss, Lisa ; Casjens, Sherwood R. ; Cingolani, Gino. / Binding-induced Stabilization and Assembly of the Phage P22 Tail Accessory Factor Gp4. In: Journal of Molecular Biology. 2006 ; Vol. 363, No. 2. pp. 558-576.
@article{98b0a375eaea45be9768753ddc25ac04,
title = "Binding-induced Stabilization and Assembly of the Phage P22 Tail Accessory Factor Gp4",
abstract = "To infect and replicate, bacteriophage P22 injects its 43 kbp genome across the cell wall of Salmonella enterica serovar Typhimurium. The attachment of phage P22 to the host cell as well as the injection of the viral DNA into the host is mediated by the virion's tail complex. This 2.8 MDa molecular machine is formed by five proteins, which include the portal protein gp1, the adhesion tailspike protein gp9, and three tail accessory factors: gp4, gp10, gp26. We have isolated the tail accessory factor gp4 and characterized its structure and binding interactions with portal protein. Interestingly, gp4 exists in solution as a monomer, which displays an exceedingly low structural stability (Tm 34 °C). Unfolded gp4 is prone to aggregation within a narrow range of temperatures both in vitro and in Salmonella extracts. In the virion the thermal unfolding of gp4 is prevented by the interaction with the dodecameric portal protein, which stabilizes the structure of gp4 and suppresses unfolded gp4 from irreversibly aggregating in the Salmonella milieu. The structural stabilization of gp4 is accompanied by the concomitant oligomerization of the protein to form a ring of 12 subunits bound to the lower end of the portal ring. The interaction of gp4 with portal protein is complex and likely involves the distinct binding of two non-equivalent sets of six gp4 proteins. Binding of the first set of six gp4 equivalents to dodecameric portal protein yields a gp(1)12:gp(4)6 assembly intermediate, which is stably populated at 30 °C and can be resolved by native gel electrophoresis. The final product of the assembly reaction is a bi-dodecameric gp(1)12:gp(4)12 complex, which appears hollow by electron microscopy, suggesting that gp4 does not physically plug the DNA entry/exit channel, but acts as a structural adaptor for the other tail accessory factors: gp10 and gp26.",
keywords = "bacteriophage P22, gp10, gp26, gp4, portal protein",
author = "Olia, {Adam S.} and Jawdat Al-Bassam and Winn-Stapley, {Danella A.} and Lisa Joss and Casjens, {Sherwood R.} and Gino Cingolani",
year = "2006",
month = "10",
day = "20",
doi = "10.1016/j.jmb.2006.08.014",
language = "English (US)",
volume = "363",
pages = "558--576",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Binding-induced Stabilization and Assembly of the Phage P22 Tail Accessory Factor Gp4

AU - Olia, Adam S.

AU - Al-Bassam, Jawdat

AU - Winn-Stapley, Danella A.

AU - Joss, Lisa

AU - Casjens, Sherwood R.

AU - Cingolani, Gino

PY - 2006/10/20

Y1 - 2006/10/20

N2 - To infect and replicate, bacteriophage P22 injects its 43 kbp genome across the cell wall of Salmonella enterica serovar Typhimurium. The attachment of phage P22 to the host cell as well as the injection of the viral DNA into the host is mediated by the virion's tail complex. This 2.8 MDa molecular machine is formed by five proteins, which include the portal protein gp1, the adhesion tailspike protein gp9, and three tail accessory factors: gp4, gp10, gp26. We have isolated the tail accessory factor gp4 and characterized its structure and binding interactions with portal protein. Interestingly, gp4 exists in solution as a monomer, which displays an exceedingly low structural stability (Tm 34 °C). Unfolded gp4 is prone to aggregation within a narrow range of temperatures both in vitro and in Salmonella extracts. In the virion the thermal unfolding of gp4 is prevented by the interaction with the dodecameric portal protein, which stabilizes the structure of gp4 and suppresses unfolded gp4 from irreversibly aggregating in the Salmonella milieu. The structural stabilization of gp4 is accompanied by the concomitant oligomerization of the protein to form a ring of 12 subunits bound to the lower end of the portal ring. The interaction of gp4 with portal protein is complex and likely involves the distinct binding of two non-equivalent sets of six gp4 proteins. Binding of the first set of six gp4 equivalents to dodecameric portal protein yields a gp(1)12:gp(4)6 assembly intermediate, which is stably populated at 30 °C and can be resolved by native gel electrophoresis. The final product of the assembly reaction is a bi-dodecameric gp(1)12:gp(4)12 complex, which appears hollow by electron microscopy, suggesting that gp4 does not physically plug the DNA entry/exit channel, but acts as a structural adaptor for the other tail accessory factors: gp10 and gp26.

AB - To infect and replicate, bacteriophage P22 injects its 43 kbp genome across the cell wall of Salmonella enterica serovar Typhimurium. The attachment of phage P22 to the host cell as well as the injection of the viral DNA into the host is mediated by the virion's tail complex. This 2.8 MDa molecular machine is formed by five proteins, which include the portal protein gp1, the adhesion tailspike protein gp9, and three tail accessory factors: gp4, gp10, gp26. We have isolated the tail accessory factor gp4 and characterized its structure and binding interactions with portal protein. Interestingly, gp4 exists in solution as a monomer, which displays an exceedingly low structural stability (Tm 34 °C). Unfolded gp4 is prone to aggregation within a narrow range of temperatures both in vitro and in Salmonella extracts. In the virion the thermal unfolding of gp4 is prevented by the interaction with the dodecameric portal protein, which stabilizes the structure of gp4 and suppresses unfolded gp4 from irreversibly aggregating in the Salmonella milieu. The structural stabilization of gp4 is accompanied by the concomitant oligomerization of the protein to form a ring of 12 subunits bound to the lower end of the portal ring. The interaction of gp4 with portal protein is complex and likely involves the distinct binding of two non-equivalent sets of six gp4 proteins. Binding of the first set of six gp4 equivalents to dodecameric portal protein yields a gp(1)12:gp(4)6 assembly intermediate, which is stably populated at 30 °C and can be resolved by native gel electrophoresis. The final product of the assembly reaction is a bi-dodecameric gp(1)12:gp(4)12 complex, which appears hollow by electron microscopy, suggesting that gp4 does not physically plug the DNA entry/exit channel, but acts as a structural adaptor for the other tail accessory factors: gp10 and gp26.

KW - bacteriophage P22

KW - gp10

KW - gp26

KW - gp4

KW - portal protein

UR - http://www.scopus.com/inward/record.url?scp=33748934964&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33748934964&partnerID=8YFLogxK

U2 - 10.1016/j.jmb.2006.08.014

DO - 10.1016/j.jmb.2006.08.014

M3 - Article

VL - 363

SP - 558

EP - 576

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 2

ER -