A combined proteomic and transcriptomic approach to the study of stage differentiation in Leishmania infantum
Protozoan parasites of the genus Leishmania are found as promastigotes in the sandfly vector and as amastigotes in mammalian macrophages. Mechanisms controlling stage‐regulated gene expression in these organisms are poorly understood. Here, we applied a comprehensive approach consisting of protein p...
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| Veröffentlicht in: | Proteomics (Weinheim) 2006-06, Vol.6 (12), p.3567-3581 |
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| creator | McNicoll, François Drummelsmith, Jolyne Müller, Michaela Madore, Éric Boilard, Nathalie Ouellette, Marc Papadopoulou, Barbara |
| description | Protozoan parasites of the genus Leishmania are found as promastigotes in the sandfly vector and as amastigotes in mammalian macrophages. Mechanisms controlling stage‐regulated gene expression in these organisms are poorly understood. Here, we applied a comprehensive approach consisting of protein prefractionation, global proteomics and targeted DNA microarray analysis to the study of stage differentiation in Leishmania. By excluding some abundant structural proteins and reducing complexity, we detected and identified numerous novel differentially expressed protein isoforms in L. infantum. Using 2‐D gels, over 2200 protein isoforms were visualized in each developmental stage. Of these, 6.1% were strongly increased or appeared unique in the promastigote stage, while the relative amounts of 12.4% were increased in amastigotes. Amastigote‐specific protein isoform and mRNA expression trends correlated modestly (53%), while no correlation was found for promastigote‐specific spots. Even where direction of regulation was similar, fold‐changes were more modest at the RNA than protein level. Many proteins were present in multiple spots, suggesting that PTM is extensive in this organism. In several cases, different isoforms appeared to be specific to different life stages. Our results suggest that post‐transcriptional controls at translational and post‐translational levels could play major roles in differentiation in Leishmania parasites. |
| doi_str_mv | 10.1002/pmic.200500853 |
| format | Article |
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Mechanisms controlling stage‐regulated gene expression in these organisms are poorly understood. Here, we applied a comprehensive approach consisting of protein prefractionation, global proteomics and targeted DNA microarray analysis to the study of stage differentiation in Leishmania. By excluding some abundant structural proteins and reducing complexity, we detected and identified numerous novel differentially expressed protein isoforms in L. infantum. Using 2‐D gels, over 2200 protein isoforms were visualized in each developmental stage. Of these, 6.1% were strongly increased or appeared unique in the promastigote stage, while the relative amounts of 12.4% were increased in amastigotes. Amastigote‐specific protein isoform and mRNA expression trends correlated modestly (53%), while no correlation was found for promastigote‐specific spots. Even where direction of regulation was similar, fold‐changes were more modest at the RNA than protein level. Many proteins were present in multiple spots, suggesting that PTM is extensive in this organism. In several cases, different isoforms appeared to be specific to different life stages. Our results suggest that post‐transcriptional controls at translational and post‐translational levels could play major roles in differentiation in Leishmania parasites.</description><identifier>ISSN: 1615-9853</identifier><identifier>EISSN: 1615-9861</identifier><identifier>DOI: 10.1002/pmic.200500853</identifier><identifier>PMID: 16705753</identifier><language>eng</language><publisher>Weinheim: WILEY-VCH Verlag</publisher><subject>Amastigote ; Analytical, structural and metabolic biochemistry ; Animals ; Biological and medical sciences ; Electrophoresis, Gel, Two-Dimensional ; Fundamental and applied biological sciences. Psychology ; Gene Expression Regulation, Developmental ; Gene Targeting ; Genes, Protozoan ; Leishmania ; Leishmania infantum ; Leishmania infantum - chemistry ; Leishmania infantum - genetics ; Leishmania infantum - growth & development ; Leishmania infantum - metabolism ; Life Cycle Stages ; Microarrays ; Miscellaneous ; Oligonucleotide Array Sequence Analysis ; Peptide Fragments - chemistry ; Peptide Mapping ; Prefractionation ; Protein Isoforms - genetics ; Protein Isoforms - metabolism ; Proteins ; Proteome - analysis ; Proteomics - methods ; Protozoan Proteins - genetics ; Protozoan Proteins - metabolism ; RNA, Messenger - metabolism ; Stage differentiation ; Transcription, Genetic</subject><ispartof>Proteomics (Weinheim), 2006-06, Vol.6 (12), p.3567-3581</ispartof><rights>Copyright © 2006 WILEY‐VCH Verlag GmbH & Co. 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Mechanisms controlling stage‐regulated gene expression in these organisms are poorly understood. Here, we applied a comprehensive approach consisting of protein prefractionation, global proteomics and targeted DNA microarray analysis to the study of stage differentiation in Leishmania. By excluding some abundant structural proteins and reducing complexity, we detected and identified numerous novel differentially expressed protein isoforms in L. infantum. Using 2‐D gels, over 2200 protein isoforms were visualized in each developmental stage. Of these, 6.1% were strongly increased or appeared unique in the promastigote stage, while the relative amounts of 12.4% were increased in amastigotes. Amastigote‐specific protein isoform and mRNA expression trends correlated modestly (53%), while no correlation was found for promastigote‐specific spots. Even where direction of regulation was similar, fold‐changes were more modest at the RNA than protein level. Many proteins were present in multiple spots, suggesting that PTM is extensive in this organism. In several cases, different isoforms appeared to be specific to different life stages. Our results suggest that post‐transcriptional controls at translational and post‐translational levels could play major roles in differentiation in Leishmania parasites.</description><subject>Amastigote</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Electrophoresis, Gel, Two-Dimensional</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression Regulation, Developmental</subject><subject>Gene Targeting</subject><subject>Genes, Protozoan</subject><subject>Leishmania</subject><subject>Leishmania infantum</subject><subject>Leishmania infantum - chemistry</subject><subject>Leishmania infantum - genetics</subject><subject>Leishmania infantum - growth & development</subject><subject>Leishmania infantum - metabolism</subject><subject>Life Cycle Stages</subject><subject>Microarrays</subject><subject>Miscellaneous</subject><subject>Oligonucleotide Array Sequence Analysis</subject><subject>Peptide Fragments - chemistry</subject><subject>Peptide Mapping</subject><subject>Prefractionation</subject><subject>Protein Isoforms - genetics</subject><subject>Protein Isoforms - metabolism</subject><subject>Proteins</subject><subject>Proteome - analysis</subject><subject>Proteomics - methods</subject><subject>Protozoan Proteins - genetics</subject><subject>Protozoan Proteins - metabolism</subject><subject>RNA, Messenger - metabolism</subject><subject>Stage differentiation</subject><subject>Transcription, Genetic</subject><issn>1615-9853</issn><issn>1615-9861</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkctv1DAQxi1ERR9w5Yh8gVuKH-vXsVpBWymlPRRV4mI58YQ1JE6IHcH-93i1y_bIyfN5ft9oNB9Cbym5pISwj9MQ2ktGiCBEC_4CnVFJRWW0pC-PteCn6DylH4RQpY16hU6pVEQowc_QcIXbcWhCBI-necwwloHYRY_z7GJq5zDl_ddU2q7d4DzivAGc8uK3eOxK4b4D9qHrYIaYg8thjDhEXENIm8HF4IrqXMzL8BqddK5P8ObwXqCvnz89rm-q-v76dn1VV4ELzSuqVoqpxsiWeO47KflqxUAYrb0qUnnqNTWM-UYYEA0w4WgLjRFM-sbQhl-gD_u5ZedfC6Rsh5Ba6HsXYVySlZooylfsvyA1RhopVAHfHcClGcDbaQ6Dm7f23yUL8P4AuNS6vivXa0N65pSWxpgdZ_bc79DD9rlP7C5Pu8vTHvO0D3e366Mq3mrvDSnDn6PXzT-tVFwJ-_Tl2mp-91R_q7U1_C80_KPq</recordid><startdate>20060601</startdate><enddate>20060601</enddate><creator>McNicoll, François</creator><creator>Drummelsmith, Jolyne</creator><creator>Müller, Michaela</creator><creator>Madore, Éric</creator><creator>Boilard, Nathalie</creator><creator>Ouellette, Marc</creator><creator>Papadopoulou, Barbara</creator><general>WILEY-VCH Verlag</general><general>WILEY‐VCH Verlag</general><general>Wiley-VCH</general><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20060601</creationdate><title>A combined proteomic and transcriptomic approach to the study of stage differentiation in Leishmania infantum</title><author>McNicoll, François ; Drummelsmith, Jolyne ; Müller, Michaela ; Madore, Éric ; Boilard, Nathalie ; Ouellette, Marc ; Papadopoulou, Barbara</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-i3583-174727b96c0d3df663442e5988d7df67d1d81922db59e5be25a1ceb9526db91b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Amastigote</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Electrophoresis, Gel, Two-Dimensional</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression Regulation, Developmental</topic><topic>Gene Targeting</topic><topic>Genes, Protozoan</topic><topic>Leishmania</topic><topic>Leishmania infantum</topic><topic>Leishmania infantum - chemistry</topic><topic>Leishmania infantum - genetics</topic><topic>Leishmania infantum - growth & development</topic><topic>Leishmania infantum - metabolism</topic><topic>Life Cycle Stages</topic><topic>Microarrays</topic><topic>Miscellaneous</topic><topic>Oligonucleotide Array Sequence Analysis</topic><topic>Peptide Fragments - chemistry</topic><topic>Peptide Mapping</topic><topic>Prefractionation</topic><topic>Protein Isoforms - genetics</topic><topic>Protein Isoforms - metabolism</topic><topic>Proteins</topic><topic>Proteome - analysis</topic><topic>Proteomics - methods</topic><topic>Protozoan Proteins - genetics</topic><topic>Protozoan Proteins - metabolism</topic><topic>RNA, Messenger - metabolism</topic><topic>Stage differentiation</topic><topic>Transcription, Genetic</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>McNicoll, François</creatorcontrib><creatorcontrib>Drummelsmith, Jolyne</creatorcontrib><creatorcontrib>Müller, Michaela</creatorcontrib><creatorcontrib>Madore, Éric</creatorcontrib><creatorcontrib>Boilard, Nathalie</creatorcontrib><creatorcontrib>Ouellette, Marc</creatorcontrib><creatorcontrib>Papadopoulou, Barbara</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Proteomics (Weinheim)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>McNicoll, François</au><au>Drummelsmith, Jolyne</au><au>Müller, Michaela</au><au>Madore, Éric</au><au>Boilard, Nathalie</au><au>Ouellette, Marc</au><au>Papadopoulou, Barbara</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A combined proteomic and transcriptomic approach to the study of stage differentiation in Leishmania infantum</atitle><jtitle>Proteomics (Weinheim)</jtitle><addtitle>Proteomics</addtitle><date>2006-06-01</date><risdate>2006</risdate><volume>6</volume><issue>12</issue><spage>3567</spage><epage>3581</epage><pages>3567-3581</pages><issn>1615-9853</issn><eissn>1615-9861</eissn><abstract>Protozoan parasites of the genus Leishmania are found as promastigotes in the sandfly vector and as amastigotes in mammalian macrophages. Mechanisms controlling stage‐regulated gene expression in these organisms are poorly understood. Here, we applied a comprehensive approach consisting of protein prefractionation, global proteomics and targeted DNA microarray analysis to the study of stage differentiation in Leishmania. By excluding some abundant structural proteins and reducing complexity, we detected and identified numerous novel differentially expressed protein isoforms in L. infantum. Using 2‐D gels, over 2200 protein isoforms were visualized in each developmental stage. Of these, 6.1% were strongly increased or appeared unique in the promastigote stage, while the relative amounts of 12.4% were increased in amastigotes. Amastigote‐specific protein isoform and mRNA expression trends correlated modestly (53%), while no correlation was found for promastigote‐specific spots. Even where direction of regulation was similar, fold‐changes were more modest at the RNA than protein level. Many proteins were present in multiple spots, suggesting that PTM is extensive in this organism. In several cases, different isoforms appeared to be specific to different life stages. Our results suggest that post‐transcriptional controls at translational and post‐translational levels could play major roles in differentiation in Leishmania parasites.</abstract><cop>Weinheim</cop><pub>WILEY-VCH Verlag</pub><pmid>16705753</pmid><doi>10.1002/pmic.200500853</doi><tpages>15</tpages></addata></record> |
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| ispartof | Proteomics (Weinheim), 2006-06, Vol.6 (12), p.3567-3581 |
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| subjects | Amastigote Analytical, structural and metabolic biochemistry Animals Biological and medical sciences Electrophoresis, Gel, Two-Dimensional Fundamental and applied biological sciences. Psychology Gene Expression Regulation, Developmental Gene Targeting Genes, Protozoan Leishmania Leishmania infantum Leishmania infantum - chemistry Leishmania infantum - genetics Leishmania infantum - growth & development Leishmania infantum - metabolism Life Cycle Stages Microarrays Miscellaneous Oligonucleotide Array Sequence Analysis Peptide Fragments - chemistry Peptide Mapping Prefractionation Protein Isoforms - genetics Protein Isoforms - metabolism Proteins Proteome - analysis Proteomics - methods Protozoan Proteins - genetics Protozoan Proteins - metabolism RNA, Messenger - metabolism Stage differentiation Transcription, Genetic |
| title | A combined proteomic and transcriptomic approach to the study of stage differentiation in Leishmania infantum |
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