Stabilization of the beta-hairpin in Mason-Pfizer monkey virus
capsid protein- a critical step for infectivity

J 2014

Stabilization of the beta-hairpin in Mason-Pfizer monkey virus capsid protein- a critical step for infectivity

OBR, Martin, Romana HADRAVOVÁ, Michal DOLEŽAL, Ivana KŘÍŽOVÁ, Veronika PAPOUŠKOVÁ et. al.

Basic information

Original name

Stabilization of the beta-hairpin in Mason-Pfizer monkey virus capsid protein- a critical step for infectivity

Authors

OBR, Martin (203 Czech Republic), Romana HADRAVOVÁ (203 Czech Republic), Michal DOLEŽAL (203 Czech Republic), Ivana KŘÍŽOVÁ (203 Czech Republic), Veronika PAPOUŠKOVÁ (203 Czech Republic, belonging to the institution), Lukáš ŽÍDEK (203 Czech Republic, guarantor, belonging to the institution), Richard HRABAL (203 Czech Republic), Tomáš RUML (203 Czech Republic) and Michaela RUMLOVÁ (203 Czech Republic)

Edition

Retrovirology, London, Biomed Central LTD, 2014, 1742-4690

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

United Kingdom of Great Britain and Northern Ireland

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 4.185

RIV identification code

RIV/00216224:14740/14:00077815

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.1186/s12977-014-0094-8

UT WoS

000344571100001

Keywords in English

N-TERMINAL DOMAIN; IN-VITRO; STRUCTURAL-ANALYSIS; ESCHERICHIA-COLI; GAG POLYPROTEIN; CORE FORMATION; RESOLUTION; MATURATION; ASSIGNMENT; MUTATIONS

Abstract

V originále

Background: Formation of a mature core is a crucial event for infectivity of retroviruses such as Mason-Pfizer monkey virus (M-PMV). The process is triggered by proteolytic cleavage of the polyprotein precursor Gag, which releases matrix, capsid (CA), and nucleocapsid proteins. Once released, CA assembles to form a mature core - a hexameric lattice protein shell that protects retroviral genomic RNA. Subtle conformational changes within CA induce the transition from the immature lattice to the mature lattice. Upon release from the precursor, the initially unstructured N-terminus of CA is refolded to form a beta-hairpin stabilized by a salt bridge between the N-terminal proline and conserved aspartate. Although the crucial role of the beta-hairpin in the mature core assembly has been confirmed, its precise structural function remains poorly understood. Results: Based on a previous NMR analysis of the N-terminal part of M-PMV CA, which suggested the role of additional interactions besides the proline-aspartate salt bridge in stabilization of the beta-hairpin, we introduced a series of mutations into the CA sequence. The effect of the mutations on virus assembly and infectivity was analyzed. In addition, the structural consequences of selected mutations were determined by NMR spectroscopy. We identified a network of interactions critical for proper formation of the M-PMV core. This network involves residue R14, located in the N-terminal beta-hairpin; residue W52 in the loop connecting helices 2 and 3; and residues Q113, Q115, and Y116 in helix 5. Conclusion: Combining functional and structural analyses, we identified a network of supportive interactions that stabilize the beta-hairpin in mature M-PMV CA.

Links

ED1.1.00/02.0068, research and development project

Name: CEITEC - central european institute of technology

261863, interní kód MU

Name: BioNMR Facilities - Bio NMR (Acronym: BioNMR)

Investor: European Union, Capacities

Citovat

OBR, Martin, Romana HADRAVOVÁ, Michal DOLEŽAL, Ivana KŘÍŽOVÁ, Veronika PAPOUŠKOVÁ, Lukáš ŽÍDEK, Richard HRABAL, Tomáš RUML and Michaela RUMLOVÁ. Stabilization of the beta-hairpin in Mason-Pfizer monkey virus capsid protein- a critical step for infectivity. Retrovirology. London: Biomed Central LTD, 2014, vol. 11, October 2014, p. "nestránkováno", 14 pp. ISSN 1742-4690. Available from: https://dx.doi.org/10.1186/s12977-014-0094-8.

@article{1213591,
   author = {Obr, Martin and Hadravová, Romana and Doležal, Michal and Křížová, Ivana and Papoušková, Veronika and Žídek, Lukáš and Hrabal, Richard and Ruml, Tomáš and Rumlová, Michaela},
   article_location = {London},
   article_number = {October 2014},
   doi = {http://dx.doi.org/10.1186/s12977-014-0094-8},
   keywords = {N-TERMINAL DOMAIN; IN-VITRO; STRUCTURAL-ANALYSIS; ESCHERICHIA-COLI; GAG POLYPROTEIN; CORE FORMATION; RESOLUTION; MATURATION; ASSIGNMENT; MUTATIONS},
   language = {eng},
   issn = {1742-4690},
   journal = {Retrovirology},
   title = {Stabilization of the beta-hairpin in Mason-Pfizer monkey virus capsid protein- a critical step for infectivity},
   url = {http://www.retrovirology.com/content/11/1/94},
   volume = {11},
   year = {2014}
}

TY  - JOUR
ID  - 1213591
AU  - Obr, Martin - Hadravová, Romana - Doležal, Michal - Křížová, Ivana - Papoušková, Veronika - Žídek, Lukáš - Hrabal, Richard - Ruml, Tomáš - Rumlová, Michaela
PY  - 2014
TI  - Stabilization of the beta-hairpin in Mason-Pfizer monkey virus capsid protein- a critical step for infectivity
JF  - Retrovirology
VL  - 11
IS  - October 2014
SP  - "nestránkováno"
EP  - "nestránkováno"
PB  - Biomed Central LTD
SN  - 17424690
KW  - N-TERMINAL DOMAIN
KW  - IN-VITRO
KW  - STRUCTURAL-ANALYSIS
KW  - ESCHERICHIA-COLI
KW  - GAG POLYPROTEIN
KW  - CORE FORMATION
KW  - RESOLUTION
KW  - MATURATION
KW  - ASSIGNMENT
KW  - MUTATIONS
UR  - http://www.retrovirology.com/content/11/1/94
L2  - http://www.retrovirology.com/content/11/1/94
N2  - Background: Formation of a mature core is a crucial event for infectivity of retroviruses such as Mason-Pfizer monkey virus (M-PMV). The process is triggered by proteolytic cleavage of the polyprotein precursor Gag, which releases matrix, capsid (CA), and nucleocapsid proteins. Once released, CA assembles to form a mature core - a hexameric lattice protein shell that protects retroviral genomic RNA. Subtle conformational changes within CA induce the transition from the immature lattice to the mature lattice. Upon release from the precursor, the initially unstructured N-terminus of CA is refolded to form a beta-hairpin stabilized by a salt bridge between the N-terminal proline and conserved aspartate. Although the crucial role of the beta-hairpin in the mature core assembly has been confirmed, its precise structural function remains poorly understood. Results: Based on a previous NMR analysis of the N-terminal part of M-PMV CA, which suggested the role of additional interactions besides the proline-aspartate salt bridge in stabilization of the beta-hairpin, we introduced a series of mutations into the CA sequence. The effect of the mutations on virus assembly and infectivity was analyzed. In addition, the structural consequences of selected mutations were determined by NMR spectroscopy. We identified a network of interactions critical for proper formation of the M-PMV core. This network involves residue R14, located in the N-terminal beta-hairpin; residue W52 in the loop connecting helices 2 and 3; and residues Q113, Q115, and Y116 in helix 5. Conclusion: Combining functional and structural analyses, we identified a network of supportive interactions that stabilize the beta-hairpin in mature M-PMV CA.
ER  -

OBR, Martin, Romana HADRAVOVÁ, Michal DOLEŽAL, Ivana KŘÍŽOVÁ, Veronika PAPOUŠKOVÁ, Lukáš ŽÍDEK, Richard HRABAL, Tomáš RUML and Michaela RUMLOVÁ. Stabilization of the beta-hairpin in Mason-Pfizer monkey virus capsid protein- a critical step for infectivity. \textit{Retrovirology}. London: Biomed Central LTD, 2014, vol.~11, October 2014, p.~''nestránkováno'', 14 pp. ISSN~1742-4690. Available from: https://dx.doi.org/10.1186/s12977-014-0094-8.

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