Molecular characterization of a novel amastigote stage specific Class I nuclease from Leishmania major
Leishmania parasites like other kinetoplastids are unable to synthesize purines de novo and so are reliant on a salvage pathway for recycling ribonucleotides. A stage specific class one nuclease enzyme, 3′-Nucleotidase/nuclease, has been implicated in salvage of preformed purines in Leishmania insec...
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Veröffentlicht in: | International journal for parasitology 2004-07, Vol.34 (8), p.899-908 |
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description | Leishmania parasites like other kinetoplastids are unable to synthesize purines de novo and so are reliant on a salvage pathway for recycling ribonucleotides. A stage specific class one nuclease enzyme, 3′-Nucleotidase/nuclease, has been implicated in salvage of preformed purines in
Leishmania insect stage promastigote via hydrolysis of 3′-nucleotides and nucleic acids. Although a similar activity is known to exist in amastigotes which reside in infected mammalian cells, the homologous gene and the corresponding protein responsible for carrying out this function have not been well characterized. Using primers specific for conserved regions of trypanosomatid class one nucleases, a gene encoding a novel class one nuclease from amastigotes of
Leishmania major (
LmaC1N) was cloned and sequenced. The coding sequence consists of 951 bp encoding a 316 amino acid protein with a predicted molecular mass of 35,300
Da. Analysis of the deduced amino acid sequence showed that
LmaC1N is highly homologous to other class I nucleases and contains all five conserved regions reported for promastigotes 3′-Nucleotidase/nuclease. Analysis by reverse transcriptase polymerase chain reaction and Western blotting demonstrated that expression of
LmaC1N gene is regulated in a stage-specific manner. Whereas the gene appeared to be silenced in promastigotes, high level expression in amastigotes implied an important function in support of parasite survival and multiplication in the mammalian cells. |
doi_str_mv | 10.1016/j.ijpara.2004.03.005 |
format | Article |
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Leishmania insect stage promastigote via hydrolysis of 3′-nucleotides and nucleic acids. Although a similar activity is known to exist in amastigotes which reside in infected mammalian cells, the homologous gene and the corresponding protein responsible for carrying out this function have not been well characterized. Using primers specific for conserved regions of trypanosomatid class one nucleases, a gene encoding a novel class one nuclease from amastigotes of
Leishmania major (
LmaC1N) was cloned and sequenced. The coding sequence consists of 951 bp encoding a 316 amino acid protein with a predicted molecular mass of 35,300
Da. Analysis of the deduced amino acid sequence showed that
LmaC1N is highly homologous to other class I nucleases and contains all five conserved regions reported for promastigotes 3′-Nucleotidase/nuclease. Analysis by reverse transcriptase polymerase chain reaction and Western blotting demonstrated that expression of
LmaC1N gene is regulated in a stage-specific manner. Whereas the gene appeared to be silenced in promastigotes, high level expression in amastigotes implied an important function in support of parasite survival and multiplication in the mammalian cells.</description><identifier>ISSN: 0020-7519</identifier><identifier>EISSN: 1879-0135</identifier><identifier>DOI: 10.1016/j.ijpara.2004.03.005</identifier><identifier>PMID: 15217728</identifier><identifier>CODEN: IJPYBT</identifier><language>eng</language><publisher>Oxford: Elsevier Ltd</publisher><subject>3′-Nnucleotidase/nuclease ; Amastigote ; Amino Acid Sequence ; Animals ; Base Sequence ; Biological and medical sciences ; Blotting, Southern - methods ; Class I nuclease ; Cloning, Molecular - methods ; Culture Media ; Fundamental and applied biological sciences. Psychology ; Gene Expression Regulation - genetics ; Leishmania major ; Leishmania major - enzymology ; Leishmania major - genetics ; Life cycle. Host-agent relationship. Pathogenesis ; Molecular Sequence Data ; Nucleotidases - genetics ; Nucleotidases - metabolism ; Protozoa ; Protozoan Proteins - genetics ; Purine metabolism ; Purine salvage ; Purines - metabolism ; Recombinant Proteins - genetics ; Reverse Transcriptase Polymerase Chain Reaction - methods ; Sequence Alignment</subject><ispartof>International journal for parasitology, 2004-07, Vol.34 (8), p.899-908</ispartof><rights>2004 Australian Society for Parasitology Inc</rights><rights>2004 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c419t-b0ecf781d2d7c592260c038c998f2d31df92410f7c6e7ed92a32cf5a244cc5e63</citedby><cites>FETCH-LOGICAL-c419t-b0ecf781d2d7c592260c038c998f2d31df92410f7c6e7ed92a32cf5a244cc5e63</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.ijpara.2004.03.005$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=15928479$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15217728$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Farajnia, S</creatorcontrib><creatorcontrib>Alimohammadian, M.H</creatorcontrib><creatorcontrib>Reiner, N.E</creatorcontrib><creatorcontrib>Karimi, M</creatorcontrib><creatorcontrib>Ajdari, S</creatorcontrib><creatorcontrib>Mahboudi, F</creatorcontrib><title>Molecular characterization of a novel amastigote stage specific Class I nuclease from Leishmania major</title><title>International journal for parasitology</title><addtitle>Int J Parasitol</addtitle><description>Leishmania parasites like other kinetoplastids are unable to synthesize purines de novo and so are reliant on a salvage pathway for recycling ribonucleotides. A stage specific class one nuclease enzyme, 3′-Nucleotidase/nuclease, has been implicated in salvage of preformed purines in
Leishmania insect stage promastigote via hydrolysis of 3′-nucleotides and nucleic acids. Although a similar activity is known to exist in amastigotes which reside in infected mammalian cells, the homologous gene and the corresponding protein responsible for carrying out this function have not been well characterized. Using primers specific for conserved regions of trypanosomatid class one nucleases, a gene encoding a novel class one nuclease from amastigotes of
Leishmania major (
LmaC1N) was cloned and sequenced. The coding sequence consists of 951 bp encoding a 316 amino acid protein with a predicted molecular mass of 35,300
Da. Analysis of the deduced amino acid sequence showed that
LmaC1N is highly homologous to other class I nucleases and contains all five conserved regions reported for promastigotes 3′-Nucleotidase/nuclease. Analysis by reverse transcriptase polymerase chain reaction and Western blotting demonstrated that expression of
LmaC1N gene is regulated in a stage-specific manner. Whereas the gene appeared to be silenced in promastigotes, high level expression in amastigotes implied an important function in support of parasite survival and multiplication in the mammalian cells.</description><subject>3′-Nnucleotidase/nuclease</subject><subject>Amastigote</subject><subject>Amino Acid Sequence</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Blotting, Southern - methods</subject><subject>Class I nuclease</subject><subject>Cloning, Molecular - methods</subject><subject>Culture Media</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Expression Regulation - genetics</subject><subject>Leishmania major</subject><subject>Leishmania major - enzymology</subject><subject>Leishmania major - genetics</subject><subject>Life cycle. Host-agent relationship. Pathogenesis</subject><subject>Molecular Sequence Data</subject><subject>Nucleotidases - genetics</subject><subject>Nucleotidases - metabolism</subject><subject>Protozoa</subject><subject>Protozoan Proteins - genetics</subject><subject>Purine metabolism</subject><subject>Purine salvage</subject><subject>Purines - metabolism</subject><subject>Recombinant Proteins - genetics</subject><subject>Reverse Transcriptase Polymerase Chain Reaction - methods</subject><subject>Sequence Alignment</subject><issn>0020-7519</issn><issn>1879-0135</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU1v1DAQhi0EokvhHyDkC9wSxo7z4QsSWgGttIgLnK3pZNw6SuLFTirBryfVrgSncpm5PO-r0TxCvFZQKlDN-6EMwxETlhrAlFCVAPUTsVNdawtQVf1U7AA0FG2t7IV4kfMAoOrKmOfiQtVata3udsJ_jSPTOmKSdLe10cIp_MYlxFlGL1HO8Z5HiRPmJdzGhWVe8HabR6bgA8n9iDnLazmvNDJmlj7FSR445LsJ54BywiGml-KZxzHzq_O-FD8-f_q-vyoO375c7z8eCjLKLsUNMPm2U73uW6qt1g0QVB1Z23ndV6r3VhsFvqWGW-6txkqTr1EbQ1RzU12Kd6feY4o_V86Lm0ImHkecOa7ZNU1T686a_4KqA6NtozbQnEBKMefE3h1TmDD9cgrcgwg3uJMI9yDCQeU2EVvszbl_vZm4_xs6f34D3p4BzISjTzhTyP9wVnemtRv34cTx9rb7wMllCjwT9yExLa6P4fFL_gD84Kky</recordid><startdate>20040701</startdate><enddate>20040701</enddate><creator>Farajnia, S</creator><creator>Alimohammadian, M.H</creator><creator>Reiner, N.E</creator><creator>Karimi, M</creator><creator>Ajdari, S</creator><creator>Mahboudi, F</creator><general>Elsevier Ltd</general><general>Elsevier Science</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>M7N</scope><scope>7X8</scope></search><sort><creationdate>20040701</creationdate><title>Molecular characterization of a novel amastigote stage specific Class I nuclease from Leishmania major</title><author>Farajnia, S ; Alimohammadian, M.H ; Reiner, N.E ; Karimi, M ; Ajdari, S ; Mahboudi, F</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c419t-b0ecf781d2d7c592260c038c998f2d31df92410f7c6e7ed92a32cf5a244cc5e63</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>3′-Nnucleotidase/nuclease</topic><topic>Amastigote</topic><topic>Amino Acid Sequence</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Blotting, Southern - methods</topic><topic>Class I nuclease</topic><topic>Cloning, Molecular - methods</topic><topic>Culture Media</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Expression Regulation - genetics</topic><topic>Leishmania major</topic><topic>Leishmania major - enzymology</topic><topic>Leishmania major - genetics</topic><topic>Life cycle. Host-agent relationship. Pathogenesis</topic><topic>Molecular Sequence Data</topic><topic>Nucleotidases - genetics</topic><topic>Nucleotidases - metabolism</topic><topic>Protozoa</topic><topic>Protozoan Proteins - genetics</topic><topic>Purine metabolism</topic><topic>Purine salvage</topic><topic>Purines - metabolism</topic><topic>Recombinant Proteins - genetics</topic><topic>Reverse Transcriptase Polymerase Chain Reaction - methods</topic><topic>Sequence Alignment</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Farajnia, S</creatorcontrib><creatorcontrib>Alimohammadian, M.H</creatorcontrib><creatorcontrib>Reiner, N.E</creatorcontrib><creatorcontrib>Karimi, M</creatorcontrib><creatorcontrib>Ajdari, S</creatorcontrib><creatorcontrib>Mahboudi, F</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>MEDLINE - Academic</collection><jtitle>International journal for parasitology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Farajnia, S</au><au>Alimohammadian, M.H</au><au>Reiner, N.E</au><au>Karimi, M</au><au>Ajdari, S</au><au>Mahboudi, F</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Molecular characterization of a novel amastigote stage specific Class I nuclease from Leishmania major</atitle><jtitle>International journal for parasitology</jtitle><addtitle>Int J Parasitol</addtitle><date>2004-07-01</date><risdate>2004</risdate><volume>34</volume><issue>8</issue><spage>899</spage><epage>908</epage><pages>899-908</pages><issn>0020-7519</issn><eissn>1879-0135</eissn><coden>IJPYBT</coden><abstract>Leishmania parasites like other kinetoplastids are unable to synthesize purines de novo and so are reliant on a salvage pathway for recycling ribonucleotides. A stage specific class one nuclease enzyme, 3′-Nucleotidase/nuclease, has been implicated in salvage of preformed purines in
Leishmania insect stage promastigote via hydrolysis of 3′-nucleotides and nucleic acids. Although a similar activity is known to exist in amastigotes which reside in infected mammalian cells, the homologous gene and the corresponding protein responsible for carrying out this function have not been well characterized. Using primers specific for conserved regions of trypanosomatid class one nucleases, a gene encoding a novel class one nuclease from amastigotes of
Leishmania major (
LmaC1N) was cloned and sequenced. The coding sequence consists of 951 bp encoding a 316 amino acid protein with a predicted molecular mass of 35,300
Da. Analysis of the deduced amino acid sequence showed that
LmaC1N is highly homologous to other class I nucleases and contains all five conserved regions reported for promastigotes 3′-Nucleotidase/nuclease. Analysis by reverse transcriptase polymerase chain reaction and Western blotting demonstrated that expression of
LmaC1N gene is regulated in a stage-specific manner. Whereas the gene appeared to be silenced in promastigotes, high level expression in amastigotes implied an important function in support of parasite survival and multiplication in the mammalian cells.</abstract><cop>Oxford</cop><pub>Elsevier Ltd</pub><pmid>15217728</pmid><doi>10.1016/j.ijpara.2004.03.005</doi><tpages>10</tpages></addata></record> |
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subjects | 3′-Nnucleotidase/nuclease Amastigote Amino Acid Sequence Animals Base Sequence Biological and medical sciences Blotting, Southern - methods Class I nuclease Cloning, Molecular - methods Culture Media Fundamental and applied biological sciences. Psychology Gene Expression Regulation - genetics Leishmania major Leishmania major - enzymology Leishmania major - genetics Life cycle. Host-agent relationship. Pathogenesis Molecular Sequence Data Nucleotidases - genetics Nucleotidases - metabolism Protozoa Protozoan Proteins - genetics Purine metabolism Purine salvage Purines - metabolism Recombinant Proteins - genetics Reverse Transcriptase Polymerase Chain Reaction - methods Sequence Alignment |
title | Molecular characterization of a novel amastigote stage specific Class I nuclease from Leishmania major |
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