Crystal structure of ORF12 from Lactococcus lactis phage p2 identifies a tape measure protein chaperone.
Identifieur interne : 002807 ( Main/Exploration ); précédent : 002806; suivant : 002808Crystal structure of ORF12 from Lactococcus lactis phage p2 identifies a tape measure protein chaperone.
Auteurs : Marina Siponen [France] ; Giuliano Sciara ; Manuela Villion ; Silvia Spinelli ; Julie Lichière ; Christian Cambillau ; Sylvain Moineau ; Valérie CampanacciSource :
- Journal of bacteriology [ 1098-5530 ] ; 2009.
Descripteurs français
- KwdFr :
- MESH :
- génétique : Bactériophage P2, Protéines virales.
- métabolisme : Bactériophage P2, Protéines virales.
- virologie : Lactococcus lactis.
- Clonage moléculaire, Cristallographie aux rayons X, Modèles moléculaires, Protéines virales, Structure secondaire des protéines.
English descriptors
- KwdEn :
- MESH :
- chemical , chemistry : Viral Proteins.
- genetics : Bacteriophage P2, Viral Proteins.
- metabolism : Bacteriophage P2, Viral Proteins.
- virology : Lactococcus lactis.
- Cloning, Molecular, Crystallography, X-Ray, Models, Molecular, Protein Structure, Secondary.
Abstract
We report here the characterization of the nonstructural protein ORF12 of the virulent lactococcal phage p2, which belongs to the Siphoviridae family. ORF12 was produced as a soluble protein, which forms large oligomers (6- to 15-mers) in solution. Using anti-ORF12 antibodies, we have confirmed that ORF12 is not found in the virion structure but is detected in the second half of the lytic cycle, indicating that it is a late-expressed protein. The structure of ORF12, solved by single anomalous diffraction and refined at 2.9-A resolution, revealed a previously unknown fold as well as the presence of a hydrophobic patch at its surface. Furthermore, crystal packing of ORF12 formed long spirals in which a hydrophobic, continuous crevice was identified. This crevice exhibited a repeated motif of aromatic residues, which coincided with the same repeated motif usually found in tape measure protein (TMP), predicted to form helices. A model of a complex between ORF12 and a repeated motif of the TMP of phage p2 (ORF14) was generated, in which the TMP helix fitted exquisitely in the crevice and the aromatic patches of ORF12. We suggest, therefore, that ORF12 might act as a chaperone for TMP hydrophobic repeats, maintaining TMP in solution during the tail assembly of the lactococcal siphophage p2.
DOI: 10.1128/JB.01363-08
PubMed: 19047351
Affiliations:
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Le document en format XML
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type="city">Marseille</settlement></placeName></affiliation></author><author><name sortKey="Sciara, Giuliano" sort="Sciara, Giuliano" uniqKey="Sciara G" first="Giuliano" last="Sciara">Giuliano Sciara</name></author><author><name sortKey="Villion, Manuela" sort="Villion, Manuela" uniqKey="Villion M" first="Manuela" last="Villion">Manuela Villion</name></author><author><name sortKey="Spinelli, Silvia" sort="Spinelli, Silvia" uniqKey="Spinelli S" first="Silvia" last="Spinelli">Silvia Spinelli</name></author><author><name sortKey="Lichiere, Julie" sort="Lichiere, Julie" uniqKey="Lichiere J" first="Julie" last="Lichière">Julie Lichière</name></author><author><name sortKey="Cambillau, Christian" sort="Cambillau, Christian" uniqKey="Cambillau C" first="Christian" last="Cambillau">Christian Cambillau</name></author><author><name sortKey="Moineau, Sylvain" sort="Moineau, Sylvain" uniqKey="Moineau S" first="Sylvain" last="Moineau">Sylvain Moineau</name></author><author><name sortKey="Campanacci, Valerie" sort="Campanacci, Valerie" uniqKey="Campanacci V" first="Valérie" last="Campanacci">Valérie Campanacci</name></author></analytic><series><title level="j">Journal of bacteriology</title><idno type="eISSN">1098-5530</idno><imprint><date when="2009" type="published">2009</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Bacteriophage P2 (genetics)</term><term>Bacteriophage P2 (metabolism)</term><term>Cloning, Molecular</term><term>Crystallography, X-Ray</term><term>Lactococcus lactis (virology)</term><term>Models, Molecular</term><term>Protein Structure, Secondary</term><term>Viral Proteins (chemistry)</term><term>Viral Proteins (genetics)</term><term>Viral Proteins (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Bactériophage P2 (génétique)</term><term>Bactériophage P2 (métabolisme)</term><term>Clonage moléculaire</term><term>Cristallographie aux rayons X</term><term>Lactococcus lactis (virologie)</term><term>Modèles moléculaires</term><term>Protéines virales ()</term><term>Protéines virales (génétique)</term><term>Protéines virales (métabolisme)</term><term>Structure secondaire des protéines</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Viral Proteins</term></keywords><keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Bacteriophage P2</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Bactériophage P2</term><term>Protéines virales</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Bacteriophage P2</term><term>Viral Proteins</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Bactériophage P2</term><term>Protéines virales</term></keywords><keywords scheme="MESH" qualifier="virologie" xml:lang="fr"><term>Lactococcus lactis</term></keywords><keywords scheme="MESH" qualifier="virology" xml:lang="en"><term>Lactococcus lactis</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Cloning, Molecular</term><term>Crystallography, X-Ray</term><term>Models, Molecular</term><term>Protein Structure, Secondary</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Clonage moléculaire</term><term>Cristallographie aux rayons X</term><term>Modèles moléculaires</term><term>Protéines virales</term><term>Structure secondaire des protéines</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">We report here the characterization of the nonstructural protein ORF12 of the virulent lactococcal phage p2, which belongs to the Siphoviridae family. ORF12 was produced as a soluble protein, which forms large oligomers (6- to 15-mers) in solution. Using anti-ORF12 antibodies, we have confirmed that ORF12 is not found in the virion structure but is detected in the second half of the lytic cycle, indicating that it is a late-expressed protein. The structure of ORF12, solved by single anomalous diffraction and refined at 2.9-A resolution, revealed a previously unknown fold as well as the presence of a hydrophobic patch at its surface. Furthermore, crystal packing of ORF12 formed long spirals in which a hydrophobic, continuous crevice was identified. This crevice exhibited a repeated motif of aromatic residues, which coincided with the same repeated motif usually found in tape measure protein (TMP), predicted to form helices. A model of a complex between ORF12 and a repeated motif of the TMP of phage p2 (ORF14) was generated, in which the TMP helix fitted exquisitely in the crevice and the aromatic patches of ORF12. We suggest, therefore, that ORF12 might act as a chaperone for TMP hydrophobic repeats, maintaining TMP in solution during the tail assembly of the lactococcal siphophage p2.</div></front></TEI><affiliations><list><country><li>France</li></country><region><li>Provence-Alpes-Côte d'Azur</li></region><settlement><li>Marseille</li></settlement></list><tree><noCountry><name sortKey="Cambillau, Christian" sort="Cambillau, Christian" uniqKey="Cambillau C" first="Christian" last="Cambillau">Christian Cambillau</name><name sortKey="Campanacci, Valerie" sort="Campanacci, Valerie" uniqKey="Campanacci V" first="Valérie" last="Campanacci">Valérie Campanacci</name><name sortKey="Lichiere, Julie" sort="Lichiere, Julie" uniqKey="Lichiere J" first="Julie" last="Lichière">Julie Lichière</name><name sortKey="Moineau, Sylvain" sort="Moineau, Sylvain" uniqKey="Moineau S" first="Sylvain" last="Moineau">Sylvain Moineau</name><name sortKey="Sciara, Giuliano" sort="Sciara, Giuliano" uniqKey="Sciara G" first="Giuliano" last="Sciara">Giuliano Sciara</name><name sortKey="Spinelli, Silvia" sort="Spinelli, Silvia" uniqKey="Spinelli S" first="Silvia" last="Spinelli">Silvia Spinelli</name><name sortKey="Villion, Manuela" sort="Villion, Manuela" uniqKey="Villion M" first="Manuela" last="Villion">Manuela Villion</name></noCountry><country name="France"><region name="Provence-Alpes-Côte d'Azur"><name sortKey="Siponen, Marina" sort="Siponen, Marina" uniqKey="Siponen M" first="Marina" last="Siponen">Marina Siponen</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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