Expression and complexity of the PRT1 multigene family of Pneumocystis carinii

1 Department of Molecular Genetics, Biochemistry & Microbiology, University of Cincinnati, Cincinnati, OH 45267-0524, USA 2 Department of Parasitology, Faculty of Pharmacy, 59006, Lille, and EA3609, Institut Pasteur de Lille, 59019, Lille, France 3 EA3609, Institut Pasteur de Lille, 59019, Lille...

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Veröffentlicht in:Microbiology (Society for General Microbiology) 2004-02, Vol.150 (2), p.293-300
Hauptverfasser: Ambrose, H. E, Keely, S. P, Aliouat, E. M, Dei-Cas, E, Wakefield, A. E, Miller, R. F, Stringer, J. R
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container_title Microbiology (Society for General Microbiology)
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creator Ambrose, H. E
Keely, S. P
Aliouat, E. M
Dei-Cas, E
Wakefield, A. E
Miller, R. F
Stringer, J. R
description 1 Department of Molecular Genetics, Biochemistry & Microbiology, University of Cincinnati, Cincinnati, OH 45267-0524, USA 2 Department of Parasitology, Faculty of Pharmacy, 59006, Lille, and EA3609, Institut Pasteur de Lille, 59019, Lille, France 3 EA3609, Institut Pasteur de Lille, 59019, Lille, and Lille-2 University Hospital, Lille, France 4 Molecular Infectious Diseases Group, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DU, UK 5 Department of Sexually Transmitted Diseases, Royal Free and University College Medical School, University College London, London WC1 6AU, UK Correspondence J. R. Stringer stringjr{at}ucmail.uc.edu Pneumocystis carinii has a multigene family, PRT1 , that encodes proteins with homology to KEX2-like proteases. PRT1 genes cluster with MSG genes near the telomeres and, like MSG, PRT1 proteins seem to be surface-expressed. The clustering of PRT1 and MSG genes suggested that expression of the two multigene families might be coordinated. Studying gene expression in P. carinii has been hampered by the lack of a culture system, and by lack of clonality in P. carinii populations in naturally infected rats, the host of this fungus. Heterogeneity can be reduced, however, by low-dose intratracheal inoculation, which can produce P. carinii populations dominated by organisms derived from a single progenitor. To study PRT1 expression, nude rats were inoculated with approximately 10 P. carinii each. The clonality of the P. carinii populations from inoculated rats was assessed by analysis of the UCS locus, a site in the genome that is known to be very heterogeneous in naturally infected rats, but nearly homogeneous in rats infected by low-dose intratracheal inoculation. Each of the populations had the same MSG gene at the UCS locus in at least 80 % of the organisms. To investigate PRT1 gene expression, RNA was amplified using primers that amplify numerous PRT1 genes. Seventy-four cloned cDNAs were sequenced, including at least 12 clones from each population of P. carinii . Many differently expressed PRT1 sequences were identified in each population, and a total of 45 different sequences were detected. However, the same PRT1 sequence was present in 15 of 74 plasmids and was found in 3 of the 5 P. carinii populations, suggesting that some PRT1 genes may be either more commonly expressed or expressed at a higher level. These data show that many members of the PRT1 gene family can be expressed in populations of P. ca
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E ; Keely, S. P ; Aliouat, E. M ; Dei-Cas, E ; Wakefield, A. E ; Miller, R. F ; Stringer, J. R</creator><creatorcontrib>Ambrose, H. E ; Keely, S. P ; Aliouat, E. M ; Dei-Cas, E ; Wakefield, A. E ; Miller, R. F ; Stringer, J. R</creatorcontrib><description>1 Department of Molecular Genetics, Biochemistry &amp; Microbiology, University of Cincinnati, Cincinnati, OH 45267-0524, USA 2 Department of Parasitology, Faculty of Pharmacy, 59006, Lille, and EA3609, Institut Pasteur de Lille, 59019, Lille, France 3 EA3609, Institut Pasteur de Lille, 59019, Lille, and Lille-2 University Hospital, Lille, France 4 Molecular Infectious Diseases Group, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DU, UK 5 Department of Sexually Transmitted Diseases, Royal Free and University College Medical School, University College London, London WC1 6AU, UK Correspondence J. R. Stringer stringjr{at}ucmail.uc.edu Pneumocystis carinii has a multigene family, PRT1 , that encodes proteins with homology to KEX2-like proteases. PRT1 genes cluster with MSG genes near the telomeres and, like MSG, PRT1 proteins seem to be surface-expressed. The clustering of PRT1 and MSG genes suggested that expression of the two multigene families might be coordinated. Studying gene expression in P. carinii has been hampered by the lack of a culture system, and by lack of clonality in P. carinii populations in naturally infected rats, the host of this fungus. Heterogeneity can be reduced, however, by low-dose intratracheal inoculation, which can produce P. carinii populations dominated by organisms derived from a single progenitor. To study PRT1 expression, nude rats were inoculated with approximately 10 P. carinii each. The clonality of the P. carinii populations from inoculated rats was assessed by analysis of the UCS locus, a site in the genome that is known to be very heterogeneous in naturally infected rats, but nearly homogeneous in rats infected by low-dose intratracheal inoculation. Each of the populations had the same MSG gene at the UCS locus in at least 80 % of the organisms. To investigate PRT1 gene expression, RNA was amplified using primers that amplify numerous PRT1 genes. Seventy-four cloned cDNAs were sequenced, including at least 12 clones from each population of P. carinii . Many differently expressed PRT1 sequences were identified in each population, and a total of 45 different sequences were detected. However, the same PRT1 sequence was present in 15 of 74 plasmids and was found in 3 of the 5 P. carinii populations, suggesting that some PRT1 genes may be either more commonly expressed or expressed at a higher level. These data show that many members of the PRT1 gene family can be expressed in populations of P. carinii derived from few progenitors and suggest that the regulation of this family is different from that governing expression of the MSG gene family. Abbreviations: MSG, major surface glycoprotein The GenBank accession numbers for the sequences reported in this paper are: MSG sequences, AY387711 – AY387716 , AY387718 and AY387720 – AY387739 ; PRT1 sequences; AY387740 – AY387742 , AY387745 – AY387763 , AY387765 , AY387766 and AY387768 – AY387784 .</description><identifier>ISSN: 1350-0872</identifier><identifier>EISSN: 1465-2080</identifier><identifier>DOI: 10.1099/mic.0.26539-0</identifier><identifier>PMID: 14766907</identifier><language>eng</language><publisher>Reading: Soc General Microbiol</publisher><subject>Animals ; Base Sequence ; Biological and medical sciences ; Disease Models, Animal ; DNA Primers ; Endopeptidases - genetics ; Fundamental and applied biological sciences. Psychology ; Fungal Proteins - genetics ; Gene Expression Regulation, Fungal - genetics ; Growth, nutrition, metabolism, transports, enzymes. Molecular biology ; Male ; Microbiology ; Molecular Sequence Data ; MSG gene ; Multigene Family ; Mycology ; Pathogenicity, host-agent relations, miscellaneous strains, epidemiology ; Pneumocystis carinii ; Pneumocystis carinii - genetics ; Pneumocystis carinii - isolation &amp; purification ; Pneumocystis Infections - microbiology ; PRT1 gene ; Rats ; RNA, Messenger - genetics ; Transcription, Genetic - genetics ; UCS gene</subject><ispartof>Microbiology (Society for General Microbiology), 2004-02, Vol.150 (2), p.293-300</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c422t-8774f7d342704f2cb9d97165d3c38e17ba7ecf0b14850793f83381a5213444f43</citedby><cites>FETCH-LOGICAL-c422t-8774f7d342704f2cb9d97165d3c38e17ba7ecf0b14850793f83381a5213444f43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&amp;idt=16126352$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14766907$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ambrose, H. E</creatorcontrib><creatorcontrib>Keely, S. P</creatorcontrib><creatorcontrib>Aliouat, E. M</creatorcontrib><creatorcontrib>Dei-Cas, E</creatorcontrib><creatorcontrib>Wakefield, A. E</creatorcontrib><creatorcontrib>Miller, R. F</creatorcontrib><creatorcontrib>Stringer, J. R</creatorcontrib><title>Expression and complexity of the PRT1 multigene family of Pneumocystis carinii</title><title>Microbiology (Society for General Microbiology)</title><addtitle>Microbiology</addtitle><description>1 Department of Molecular Genetics, Biochemistry &amp; Microbiology, University of Cincinnati, Cincinnati, OH 45267-0524, USA 2 Department of Parasitology, Faculty of Pharmacy, 59006, Lille, and EA3609, Institut Pasteur de Lille, 59019, Lille, France 3 EA3609, Institut Pasteur de Lille, 59019, Lille, and Lille-2 University Hospital, Lille, France 4 Molecular Infectious Diseases Group, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DU, UK 5 Department of Sexually Transmitted Diseases, Royal Free and University College Medical School, University College London, London WC1 6AU, UK Correspondence J. R. Stringer stringjr{at}ucmail.uc.edu Pneumocystis carinii has a multigene family, PRT1 , that encodes proteins with homology to KEX2-like proteases. PRT1 genes cluster with MSG genes near the telomeres and, like MSG, PRT1 proteins seem to be surface-expressed. The clustering of PRT1 and MSG genes suggested that expression of the two multigene families might be coordinated. Studying gene expression in P. carinii has been hampered by the lack of a culture system, and by lack of clonality in P. carinii populations in naturally infected rats, the host of this fungus. Heterogeneity can be reduced, however, by low-dose intratracheal inoculation, which can produce P. carinii populations dominated by organisms derived from a single progenitor. To study PRT1 expression, nude rats were inoculated with approximately 10 P. carinii each. The clonality of the P. carinii populations from inoculated rats was assessed by analysis of the UCS locus, a site in the genome that is known to be very heterogeneous in naturally infected rats, but nearly homogeneous in rats infected by low-dose intratracheal inoculation. Each of the populations had the same MSG gene at the UCS locus in at least 80 % of the organisms. To investigate PRT1 gene expression, RNA was amplified using primers that amplify numerous PRT1 genes. Seventy-four cloned cDNAs were sequenced, including at least 12 clones from each population of P. carinii . Many differently expressed PRT1 sequences were identified in each population, and a total of 45 different sequences were detected. However, the same PRT1 sequence was present in 15 of 74 plasmids and was found in 3 of the 5 P. carinii populations, suggesting that some PRT1 genes may be either more commonly expressed or expressed at a higher level. These data show that many members of the PRT1 gene family can be expressed in populations of P. carinii derived from few progenitors and suggest that the regulation of this family is different from that governing expression of the MSG gene family. Abbreviations: MSG, major surface glycoprotein The GenBank accession numbers for the sequences reported in this paper are: MSG sequences, AY387711 – AY387716 , AY387718 and AY387720 – AY387739 ; PRT1 sequences; AY387740 – AY387742 , AY387745 – AY387763 , AY387765 , AY387766 and AY387768 – AY387784 .</description><subject>Animals</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Disease Models, Animal</subject><subject>DNA Primers</subject><subject>Endopeptidases - genetics</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fungal Proteins - genetics</subject><subject>Gene Expression Regulation, Fungal - genetics</subject><subject>Growth, nutrition, metabolism, transports, enzymes. Molecular biology</subject><subject>Male</subject><subject>Microbiology</subject><subject>Molecular Sequence Data</subject><subject>MSG gene</subject><subject>Multigene Family</subject><subject>Mycology</subject><subject>Pathogenicity, host-agent relations, miscellaneous strains, epidemiology</subject><subject>Pneumocystis carinii</subject><subject>Pneumocystis carinii - genetics</subject><subject>Pneumocystis carinii - isolation &amp; purification</subject><subject>Pneumocystis Infections - microbiology</subject><subject>PRT1 gene</subject><subject>Rats</subject><subject>RNA, Messenger - genetics</subject><subject>Transcription, Genetic - genetics</subject><subject>UCS gene</subject><issn>1350-0872</issn><issn>1465-2080</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkM1L7DAUxYMo6lOXbqUbxU3Hm6-mWcqgvgeiIroOaZo4kbYZkxad_96MM-DywYV74fzuOXAQOsUwwyDlVe_NDGak4lSWsIMOMat4SaCG3XxTDiXUghygPym9A2QR8D46wExUlQRxiB5uvpbRpuTDUOihLUzol5398uOqCK4YF7Z4en7BRT91o3-zgy2c7n33Iz4NduqDWaXRp8Lo6Afvj9Ge012yJ9t9hF5vb17mf8v7x7t_8-v70jBCxrIWgjnRUkYEMEdMI1spcMVbamhtsWi0sMZBg1nNQUjqakprrDnBlDHmGD1CFxvfZQwfk02j6n0ytuv0YMOUVA2Yc8LhvyCWpGJ5MlhuQBNDStE6tYy-13GlMKh10_nRKFA_Tau18dnWeGp62_7S22ozcL4FdDK6c1EPxqdfrsKkopxk7nLDLfzb4tNHq3LPOSuGxod1KOY5VRFJ6Tdoo5Mg</recordid><startdate>20040201</startdate><enddate>20040201</enddate><creator>Ambrose, H. 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Molecular biology</topic><topic>Male</topic><topic>Microbiology</topic><topic>Molecular Sequence Data</topic><topic>MSG gene</topic><topic>Multigene Family</topic><topic>Mycology</topic><topic>Pathogenicity, host-agent relations, miscellaneous strains, epidemiology</topic><topic>Pneumocystis carinii</topic><topic>Pneumocystis carinii - genetics</topic><topic>Pneumocystis carinii - isolation &amp; purification</topic><topic>Pneumocystis Infections - microbiology</topic><topic>PRT1 gene</topic><topic>Rats</topic><topic>RNA, Messenger - genetics</topic><topic>Transcription, Genetic - genetics</topic><topic>UCS gene</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ambrose, H. E</creatorcontrib><creatorcontrib>Keely, S. P</creatorcontrib><creatorcontrib>Aliouat, E. M</creatorcontrib><creatorcontrib>Dei-Cas, E</creatorcontrib><creatorcontrib>Wakefield, A. E</creatorcontrib><creatorcontrib>Miller, R. F</creatorcontrib><creatorcontrib>Stringer, J. 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R</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Expression and complexity of the PRT1 multigene family of Pneumocystis carinii</atitle><jtitle>Microbiology (Society for General Microbiology)</jtitle><addtitle>Microbiology</addtitle><date>2004-02-01</date><risdate>2004</risdate><volume>150</volume><issue>2</issue><spage>293</spage><epage>300</epage><pages>293-300</pages><issn>1350-0872</issn><eissn>1465-2080</eissn><abstract>1 Department of Molecular Genetics, Biochemistry &amp; Microbiology, University of Cincinnati, Cincinnati, OH 45267-0524, USA 2 Department of Parasitology, Faculty of Pharmacy, 59006, Lille, and EA3609, Institut Pasteur de Lille, 59019, Lille, France 3 EA3609, Institut Pasteur de Lille, 59019, Lille, and Lille-2 University Hospital, Lille, France 4 Molecular Infectious Diseases Group, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford OX3 9DU, UK 5 Department of Sexually Transmitted Diseases, Royal Free and University College Medical School, University College London, London WC1 6AU, UK Correspondence J. R. Stringer stringjr{at}ucmail.uc.edu Pneumocystis carinii has a multigene family, PRT1 , that encodes proteins with homology to KEX2-like proteases. PRT1 genes cluster with MSG genes near the telomeres and, like MSG, PRT1 proteins seem to be surface-expressed. The clustering of PRT1 and MSG genes suggested that expression of the two multigene families might be coordinated. Studying gene expression in P. carinii has been hampered by the lack of a culture system, and by lack of clonality in P. carinii populations in naturally infected rats, the host of this fungus. Heterogeneity can be reduced, however, by low-dose intratracheal inoculation, which can produce P. carinii populations dominated by organisms derived from a single progenitor. To study PRT1 expression, nude rats were inoculated with approximately 10 P. carinii each. The clonality of the P. carinii populations from inoculated rats was assessed by analysis of the UCS locus, a site in the genome that is known to be very heterogeneous in naturally infected rats, but nearly homogeneous in rats infected by low-dose intratracheal inoculation. Each of the populations had the same MSG gene at the UCS locus in at least 80 % of the organisms. To investigate PRT1 gene expression, RNA was amplified using primers that amplify numerous PRT1 genes. Seventy-four cloned cDNAs were sequenced, including at least 12 clones from each population of P. carinii . Many differently expressed PRT1 sequences were identified in each population, and a total of 45 different sequences were detected. However, the same PRT1 sequence was present in 15 of 74 plasmids and was found in 3 of the 5 P. carinii populations, suggesting that some PRT1 genes may be either more commonly expressed or expressed at a higher level. These data show that many members of the PRT1 gene family can be expressed in populations of P. carinii derived from few progenitors and suggest that the regulation of this family is different from that governing expression of the MSG gene family. Abbreviations: MSG, major surface glycoprotein The GenBank accession numbers for the sequences reported in this paper are: MSG sequences, AY387711 – AY387716 , AY387718 and AY387720 – AY387739 ; PRT1 sequences; AY387740 – AY387742 , AY387745 – AY387763 , AY387765 , AY387766 and AY387768 – AY387784 .</abstract><cop>Reading</cop><pub>Soc General Microbiol</pub><pmid>14766907</pmid><doi>10.1099/mic.0.26539-0</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record>
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source MEDLINE; PubMed Central
subjects Animals
Base Sequence
Biological and medical sciences
Disease Models, Animal
DNA Primers
Endopeptidases - genetics
Fundamental and applied biological sciences. Psychology
Fungal Proteins - genetics
Gene Expression Regulation, Fungal - genetics
Growth, nutrition, metabolism, transports, enzymes. Molecular biology
Male
Microbiology
Molecular Sequence Data
MSG gene
Multigene Family
Mycology
Pathogenicity, host-agent relations, miscellaneous strains, epidemiology
Pneumocystis carinii
Pneumocystis carinii - genetics
Pneumocystis carinii - isolation & purification
Pneumocystis Infections - microbiology
PRT1 gene
Rats
RNA, Messenger - genetics
Transcription, Genetic - genetics
UCS gene
title Expression and complexity of the PRT1 multigene family of Pneumocystis carinii
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