Characterization of the protease domain of Rice tungro bacilliform virus responsible for the processing of the capsid protein from the polyprotein
<p>Abstract</p> <p>Background</p> <p>Rice tungro bacilliform virus (RTBV) is a pararetrovirus, and a member of the family <it>Caulimoviridae </it>in the genus <it>Badnavirus</it>. RTBV has a long open reading frame that encodes a large polyprotei...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2005-04-01
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| Series: | Virology Journal |
| Online Access: | http://www.virologyj.com/content/2/1/33 |
| _version_ | 1818028935182548992 |
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| author | Beachy Roger N de Kochko Alexandre Rojas-Mendoza Ana Marmey Philippe Fauquet Claude M |
| author_facet | Beachy Roger N de Kochko Alexandre Rojas-Mendoza Ana Marmey Philippe Fauquet Claude M |
| author_sort | Beachy Roger N |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Rice tungro bacilliform virus (RTBV) is a pararetrovirus, and a member of the family <it>Caulimoviridae </it>in the genus <it>Badnavirus</it>. RTBV has a long open reading frame that encodes a large polyprotein (P3). Pararetroviruses show similarities with retroviruses in molecular organization and replication. P3 contains a putative movement protein (MP), the capsid protein (CP), the aspartate protease (PR) and the reverse transcriptase (RT) with a ribonuclease H activity. PR is a member of the cluster of retroviral proteases and serves to proteolytically process P3. Previous work established the N- and C-terminal amino acid sequences of CP and RT, processing of RT by PR, and estimated the molecular mass of PR by western blot assays.</p> <p>Results</p> <p>A molecular mass of a protein that was associated with virions was determined by in-line HPLC electrospray ionization mass spectral analysis. Comparison with retroviral proteases amino acid sequences allowed the characterization of a putative protease domain in this protein. Structural modelling revealed strong resemblance with retroviral proteases, with overall folds surrounding the active site being well conserved. Expression in <it>E. coli </it>of putative domain was affected by the presence or absence of the active site in the construct. Analysis of processing of CP by PR, using pulse chase labelling experiments, demonstrated that the 37 kDa capsid protein was dependent on the presence of the protease in the constructs.</p> <p>Conclusion</p> <p>The findings suggest the characterization of the RTBV protease domain. Sequence analysis, structural modelling, <it>in vitro </it>expression studies are evidence to consider the putative domain as being the protease domain. Analysis of expression of different peptides corresponding to various domains of P3 suggests a processing of CP by PR. This work clarifies the organization of the RTBV polyprotein, and its processing by the RTBV protease.</p> |
| first_indexed | 2024-12-10T05:11:41Z |
| format | Article |
| id | doaj.art-7220f358bd4748f6a6a93a3c6fac41fa |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-12-10T05:11:41Z |
| publishDate | 2005-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj.art-7220f358bd4748f6a6a93a3c6fac41fa2022-12-22T02:01:05ZengBMCVirology Journal1743-422X2005-04-01213310.1186/1743-422X-2-33Characterization of the protease domain of Rice tungro bacilliform virus responsible for the processing of the capsid protein from the polyproteinBeachy Roger Nde Kochko AlexandreRojas-Mendoza AnaMarmey PhilippeFauquet Claude M<p>Abstract</p> <p>Background</p> <p>Rice tungro bacilliform virus (RTBV) is a pararetrovirus, and a member of the family <it>Caulimoviridae </it>in the genus <it>Badnavirus</it>. RTBV has a long open reading frame that encodes a large polyprotein (P3). Pararetroviruses show similarities with retroviruses in molecular organization and replication. P3 contains a putative movement protein (MP), the capsid protein (CP), the aspartate protease (PR) and the reverse transcriptase (RT) with a ribonuclease H activity. PR is a member of the cluster of retroviral proteases and serves to proteolytically process P3. Previous work established the N- and C-terminal amino acid sequences of CP and RT, processing of RT by PR, and estimated the molecular mass of PR by western blot assays.</p> <p>Results</p> <p>A molecular mass of a protein that was associated with virions was determined by in-line HPLC electrospray ionization mass spectral analysis. Comparison with retroviral proteases amino acid sequences allowed the characterization of a putative protease domain in this protein. Structural modelling revealed strong resemblance with retroviral proteases, with overall folds surrounding the active site being well conserved. Expression in <it>E. coli </it>of putative domain was affected by the presence or absence of the active site in the construct. Analysis of processing of CP by PR, using pulse chase labelling experiments, demonstrated that the 37 kDa capsid protein was dependent on the presence of the protease in the constructs.</p> <p>Conclusion</p> <p>The findings suggest the characterization of the RTBV protease domain. Sequence analysis, structural modelling, <it>in vitro </it>expression studies are evidence to consider the putative domain as being the protease domain. Analysis of expression of different peptides corresponding to various domains of P3 suggests a processing of CP by PR. This work clarifies the organization of the RTBV polyprotein, and its processing by the RTBV protease.</p>http://www.virologyj.com/content/2/1/33 |
| spellingShingle | Beachy Roger N de Kochko Alexandre Rojas-Mendoza Ana Marmey Philippe Fauquet Claude M Characterization of the protease domain of Rice tungro bacilliform virus responsible for the processing of the capsid protein from the polyprotein Virology Journal |
| title | Characterization of the protease domain of Rice tungro bacilliform virus responsible for the processing of the capsid protein from the polyprotein |
| title_full | Characterization of the protease domain of Rice tungro bacilliform virus responsible for the processing of the capsid protein from the polyprotein |
| title_fullStr | Characterization of the protease domain of Rice tungro bacilliform virus responsible for the processing of the capsid protein from the polyprotein |
| title_full_unstemmed | Characterization of the protease domain of Rice tungro bacilliform virus responsible for the processing of the capsid protein from the polyprotein |
| title_short | Characterization of the protease domain of Rice tungro bacilliform virus responsible for the processing of the capsid protein from the polyprotein |
| title_sort | characterization of the protease domain of rice tungro bacilliform virus responsible for the processing of the capsid protein from the polyprotein |
| url | http://www.virologyj.com/content/2/1/33 |
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