The novel repressor Rce2 competes with Ace3 to regulate cellulase gene expression in the filamentous fungus Trichoderma reesei
The filamentous fungus Trichoderma reesei is widely used for industrial cellulase production. In T. reesei, cellulase gene expression is tightly controlled by a regulatory network involving multiple transcription factors. Here, we isolated a novel protein, Rce2, using a pull‐down assay and mass spec...
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| Veröffentlicht in: | Molecular microbiology 2021-11, Vol.116 (5), p.1298-1314 |
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| creator | Chen, Yumeng Wang, Wei Liu, Pei Lin, Aibo Fan, Xingjia Wu, Chuan Li, Ni Wei, Liujing Wei, Dongzhi |
| description | The filamentous fungus Trichoderma reesei is widely used for industrial cellulase production. In T. reesei, cellulase gene expression is tightly controlled by a regulatory network involving multiple transcription factors. Here, we isolated a novel protein, Rce2, using a pull‐down assay and mass spectrometry analysis, from a partial carbon catabolite de‐repression mutant, T. reesei Rut‐C30, cultured under glucose‐repressing conditions. Deletion and overexpression of Rce2 in T. reesei wild‐type QM6a and mutant Rut‐C30 revealed that Rce2 acts as a repressor of cellulase gene expression. DNase I footprinting assays, electrophoretic mobility shift assays, and chromatin immunoprecipitation assays revealed that Rce2 was located in the nucleus and bound to the consensus sequences 5′‐(T/A)NNNNCCG‐3′ and 5′‐CGGNNNN(T/A)‐3′ in the promoters of cellulase‐related genes to repress their transcription. Additionally, Rce2 antagonized Ace3 binding to the cbh1 promoter to repress its transcription. However, Rce2 was not involved in Cre1‐mediated carbon catabolite repression. These results demonstrate the mechanism through which Rce2 represses the expression of cellulase genes and provide novel insights into the regulatory system of cellulases and methods that can be used for the regulation of gene expression in T. reesei.
We isolated a novel repressor of cellulase gene expression, Rce2, using a pull‐down assay and mass spectrometry analysis, from the filamentous fungus Trichoderma reesei Rut‐C30, cultured under glucose‐repressing conditions. Our data revealed that Rce2 antagonized the activator Ace3 binding to thecellulase cbh1 promoter to repress its transcription. Our results demonstrate the mechanism through which Rce2 represses the expression of cellulase genes and provide novel insights into the regulatory system of cellulases and methods that can be used for regulation of gene expression in T. reesei. |
| doi_str_mv | 10.1111/mmi.14825 |
| format | Article |
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We isolated a novel repressor of cellulase gene expression, Rce2, using a pull‐down assay and mass spectrometry analysis, from the filamentous fungus Trichoderma reesei Rut‐C30, cultured under glucose‐repressing conditions. Our data revealed that Rce2 antagonized the activator Ace3 binding to thecellulase cbh1 promoter to repress its transcription. Our results demonstrate the mechanism through which Rce2 represses the expression of cellulase genes and provide novel insights into the regulatory system of cellulases and methods that can be used for regulation of gene expression in T. reesei.</description><identifier>ISSN: 0950-382X</identifier><identifier>EISSN: 1365-2958</identifier><identifier>DOI: 10.1111/mmi.14825</identifier><identifier>PMID: 34608686</identifier><language>eng</language><publisher>England: Blackwell Publishing Ltd</publisher><subject>Amino Acid Sequence ; Assaying ; Carbon ; Catabolite Repression ; Cellulase ; Cellulase - biosynthesis ; Cellulase - genetics ; Chromatin ; Chromosomal Proteins, Non-Histone - genetics ; Chromosomal Proteins, Non-Histone - metabolism ; Deoxyribonuclease ; Electrophoretic mobility ; Footprinting ; Fungal Proteins - genetics ; Fungal Proteins - metabolism ; Fungi ; Gene expression ; Gene Expression Regulation, Fungal ; gene regulation ; Genes ; Hypocreales - genetics ; Hypocreales - metabolism ; Immunoprecipitation ; Industrial Microbiology ; Mass spectrometry ; Mass spectroscopy ; Mutants ; Phylogeny ; Promoter Regions, Genetic ; repressor ; Repressor Proteins - genetics ; Repressor Proteins - metabolism ; transcription factor ; Transcription factors ; Trichoderma reesei</subject><ispartof>Molecular microbiology, 2021-11, Vol.116 (5), p.1298-1314</ispartof><rights>2021 John Wiley & Sons Ltd</rights><rights>2021 John Wiley & Sons Ltd.</rights><rights>Copyright © 2021 John Wiley & Sons Ltd</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3535-1dd05263e26bdd7a28fd49660d62e1340f2d71b0e0fa89a64bb669b084d30cd13</citedby><cites>FETCH-LOGICAL-c3535-1dd05263e26bdd7a28fd49660d62e1340f2d71b0e0fa89a64bb669b084d30cd13</cites><orcidid>0000-0002-0440-0859</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fmmi.14825$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fmmi.14825$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,1427,27901,27902,45550,45551,46384,46808</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/34608686$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Chen, Yumeng</creatorcontrib><creatorcontrib>Wang, Wei</creatorcontrib><creatorcontrib>Liu, Pei</creatorcontrib><creatorcontrib>Lin, Aibo</creatorcontrib><creatorcontrib>Fan, Xingjia</creatorcontrib><creatorcontrib>Wu, Chuan</creatorcontrib><creatorcontrib>Li, Ni</creatorcontrib><creatorcontrib>Wei, Liujing</creatorcontrib><creatorcontrib>Wei, Dongzhi</creatorcontrib><title>The novel repressor Rce2 competes with Ace3 to regulate cellulase gene expression in the filamentous fungus Trichoderma reesei</title><title>Molecular microbiology</title><addtitle>Mol Microbiol</addtitle><description>The filamentous fungus Trichoderma reesei is widely used for industrial cellulase production. In T. reesei, cellulase gene expression is tightly controlled by a regulatory network involving multiple transcription factors. Here, we isolated a novel protein, Rce2, using a pull‐down assay and mass spectrometry analysis, from a partial carbon catabolite de‐repression mutant, T. reesei Rut‐C30, cultured under glucose‐repressing conditions. Deletion and overexpression of Rce2 in T. reesei wild‐type QM6a and mutant Rut‐C30 revealed that Rce2 acts as a repressor of cellulase gene expression. DNase I footprinting assays, electrophoretic mobility shift assays, and chromatin immunoprecipitation assays revealed that Rce2 was located in the nucleus and bound to the consensus sequences 5′‐(T/A)NNNNCCG‐3′ and 5′‐CGGNNNN(T/A)‐3′ in the promoters of cellulase‐related genes to repress their transcription. Additionally, Rce2 antagonized Ace3 binding to the cbh1 promoter to repress its transcription. However, Rce2 was not involved in Cre1‐mediated carbon catabolite repression. These results demonstrate the mechanism through which Rce2 represses the expression of cellulase genes and provide novel insights into the regulatory system of cellulases and methods that can be used for the regulation of gene expression in T. reesei.
We isolated a novel repressor of cellulase gene expression, Rce2, using a pull‐down assay and mass spectrometry analysis, from the filamentous fungus Trichoderma reesei Rut‐C30, cultured under glucose‐repressing conditions. Our data revealed that Rce2 antagonized the activator Ace3 binding to thecellulase cbh1 promoter to repress its transcription. Our results demonstrate the mechanism through which Rce2 represses the expression of cellulase genes and provide novel insights into the regulatory system of cellulases and methods that can be used for regulation of gene expression in T. reesei.</description><subject>Amino Acid Sequence</subject><subject>Assaying</subject><subject>Carbon</subject><subject>Catabolite Repression</subject><subject>Cellulase</subject><subject>Cellulase - biosynthesis</subject><subject>Cellulase - genetics</subject><subject>Chromatin</subject><subject>Chromosomal Proteins, Non-Histone - genetics</subject><subject>Chromosomal Proteins, Non-Histone - metabolism</subject><subject>Deoxyribonuclease</subject><subject>Electrophoretic mobility</subject><subject>Footprinting</subject><subject>Fungal Proteins - genetics</subject><subject>Fungal Proteins - metabolism</subject><subject>Fungi</subject><subject>Gene expression</subject><subject>Gene Expression Regulation, Fungal</subject><subject>gene regulation</subject><subject>Genes</subject><subject>Hypocreales - genetics</subject><subject>Hypocreales - metabolism</subject><subject>Immunoprecipitation</subject><subject>Industrial Microbiology</subject><subject>Mass spectrometry</subject><subject>Mass spectroscopy</subject><subject>Mutants</subject><subject>Phylogeny</subject><subject>Promoter Regions, Genetic</subject><subject>repressor</subject><subject>Repressor Proteins - genetics</subject><subject>Repressor Proteins - metabolism</subject><subject>transcription factor</subject><subject>Transcription factors</subject><subject>Trichoderma reesei</subject><issn>0950-382X</issn><issn>1365-2958</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp10U1LwzAYB_Agis7pwS8gAS96qOalydKjiC8DhyATvJW0ebpltM1MWl8ufnbjNj0I5vLk8MufJ_wROqLknMZz0TT2nKaKiS00oFyKhGVCbaMByQRJuGLPe2g_hAUhlBPJd9EeTyVRUskB-pzOAbfuFWrsYekhBOfxYwkMl65ZQgcBv9luji9L4LhzEc36WneAS6jreAuAZ9AChvfVY-tabFvcxdDK1rqBtnN9wFXfzuKYelvOnQHf6BgEAewB2ql0HeBwM4fo6eZ6enWX3D_cjq8u75OSCy4SagwRTHJgsjBmpJmqTJpJSYxkQHlKKmZGtCBAKq0yLdOikDIriEoNJ6WhfIhO17lL7156CF3e2PD9Bd1CXDBnYpTxkcykivTkD1243rdxu6iyyFIuWFRna1V6F4KHKl9622j_kVOSf5eSx1LyVSnRHm8S-6IB8yt_WojgYg3ebA0f_yflk8l4HfkFDqSXKw</recordid><startdate>202111</startdate><enddate>202111</enddate><creator>Chen, Yumeng</creator><creator>Wang, Wei</creator><creator>Liu, Pei</creator><creator>Lin, Aibo</creator><creator>Fan, Xingjia</creator><creator>Wu, Chuan</creator><creator>Li, Ni</creator><creator>Wei, Liujing</creator><creator>Wei, Dongzhi</creator><general>Blackwell Publishing Ltd</general><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>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0002-0440-0859</orcidid></search><sort><creationdate>202111</creationdate><title>The novel repressor Rce2 competes with Ace3 to regulate cellulase gene expression in the filamentous fungus Trichoderma reesei</title><author>Chen, Yumeng ; 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In T. reesei, cellulase gene expression is tightly controlled by a regulatory network involving multiple transcription factors. Here, we isolated a novel protein, Rce2, using a pull‐down assay and mass spectrometry analysis, from a partial carbon catabolite de‐repression mutant, T. reesei Rut‐C30, cultured under glucose‐repressing conditions. Deletion and overexpression of Rce2 in T. reesei wild‐type QM6a and mutant Rut‐C30 revealed that Rce2 acts as a repressor of cellulase gene expression. DNase I footprinting assays, electrophoretic mobility shift assays, and chromatin immunoprecipitation assays revealed that Rce2 was located in the nucleus and bound to the consensus sequences 5′‐(T/A)NNNNCCG‐3′ and 5′‐CGGNNNN(T/A)‐3′ in the promoters of cellulase‐related genes to repress their transcription. Additionally, Rce2 antagonized Ace3 binding to the cbh1 promoter to repress its transcription. However, Rce2 was not involved in Cre1‐mediated carbon catabolite repression. These results demonstrate the mechanism through which Rce2 represses the expression of cellulase genes and provide novel insights into the regulatory system of cellulases and methods that can be used for the regulation of gene expression in T. reesei.
We isolated a novel repressor of cellulase gene expression, Rce2, using a pull‐down assay and mass spectrometry analysis, from the filamentous fungus Trichoderma reesei Rut‐C30, cultured under glucose‐repressing conditions. Our data revealed that Rce2 antagonized the activator Ace3 binding to thecellulase cbh1 promoter to repress its transcription. Our results demonstrate the mechanism through which Rce2 represses the expression of cellulase genes and provide novel insights into the regulatory system of cellulases and methods that can be used for regulation of gene expression in T. reesei.</abstract><cop>England</cop><pub>Blackwell Publishing Ltd</pub><pmid>34608686</pmid><doi>10.1111/mmi.14825</doi><tpages>17</tpages><orcidid>https://orcid.org/0000-0002-0440-0859</orcidid></addata></record> |
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| subjects | Amino Acid Sequence Assaying Carbon Catabolite Repression Cellulase Cellulase - biosynthesis Cellulase - genetics Chromatin Chromosomal Proteins, Non-Histone - genetics Chromosomal Proteins, Non-Histone - metabolism Deoxyribonuclease Electrophoretic mobility Footprinting Fungal Proteins - genetics Fungal Proteins - metabolism Fungi Gene expression Gene Expression Regulation, Fungal gene regulation Genes Hypocreales - genetics Hypocreales - metabolism Immunoprecipitation Industrial Microbiology Mass spectrometry Mass spectroscopy Mutants Phylogeny Promoter Regions, Genetic repressor Repressor Proteins - genetics Repressor Proteins - metabolism transcription factor Transcription factors Trichoderma reesei |
| title | The novel repressor Rce2 competes with Ace3 to regulate cellulase gene expression in the filamentous fungus Trichoderma reesei |
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