Cloning and characterization of the Neurospora crassa cyt-5 gene. A nuclear-coded mitochondrial RNA polymerase with a polyglutamine repeat

The Neurospora crassa mutants, cyt-5-1 and cyt-5-4, have a cytochrome b- and aa3-deficient phenotype, suggesting that they result from a deficiency in a nuclear-coded component of the mitochondrial gene expression apparatus (Bertrand, H., Nargang, F. E., Collins, R. A., and Zagozeski, C. A. (1977) M...

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Veröffentlicht in:The Journal of biological chemistry 1996-03, Vol.271 (11), p.6537-6544
Hauptverfasser: Chen, B. (Ohio State University, Columbus, OH.), Kubelik, A.R, Mohr, S, Breitenberger, C.A
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container_issue 11
container_start_page 6537
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creator Chen, B. (Ohio State University, Columbus, OH.)
Kubelik, A.R
Mohr, S
Breitenberger, C.A
description The Neurospora crassa mutants, cyt-5-1 and cyt-5-4, have a cytochrome b- and aa3-deficient phenotype, suggesting that they result from a deficiency in a nuclear-coded component of the mitochondrial gene expression apparatus (Bertrand, H., Nargang, F. E., Collins, R. A., and Zagozeski, C. A. (1977) Mol. Gen. Genet. 153, 247-257). The complementing wild-type gene has been cloned and sequenced, and shown to encode a protein with significant sequence similarity to Saccharomyces cerevisiae mitochondrial RNA polymerase and bacteriophage RNA polymerases. There are remarkable differences between the N. crassa protein and its yeast homologue, including a region of very little homology near the N termini of the two gene products. The cyt-5 gene encodes a stretch of polyglutamine in this region of unique sequence. In addition, an acidic insertion (86 amino acids, of which 24 are Asp or Glu and 10 are Arg or Lys) is present near the C terminus of the cyt-5 gene product. Transcript levels of the cytochrome b and cytochrome oxidase subunit III genes are severely reduced in cyt-5 mutants, suggesting a likely mechanism for the cytochrome-deficient phenotype. In contrast, mitochondrial rRNAs accumulate to nearly normal levels in cyt-5 mutants. However, mitochondrial rRNA levels are not indicative of the rate of transcription of the corresponding genes, since crude lysates of mitochondria from cyt-5 mutants exhibit greatly reduced transcriptional activity with a 19 S rRNA promoter. The cyt-5 gene is flanked by at least one gene whose product also may be involved in mitochondrial function
doi_str_mv 10.1074/jbc.271.11.6537
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A nuclear-coded mitochondrial RNA polymerase with a polyglutamine repeat</title><source>MEDLINE</source><source>EZB-FREE-00999 freely available EZB journals</source><source>Alma/SFX Local Collection</source><creator>Chen, B. (Ohio State University, Columbus, OH.) ; Kubelik, A.R ; Mohr, S ; Breitenberger, C.A</creator><creatorcontrib>Chen, B. (Ohio State University, Columbus, OH.) ; Kubelik, A.R ; Mohr, S ; Breitenberger, C.A</creatorcontrib><description>The Neurospora crassa mutants, cyt-5-1 and cyt-5-4, have a cytochrome b- and aa3-deficient phenotype, suggesting that they result from a deficiency in a nuclear-coded component of the mitochondrial gene expression apparatus (Bertrand, H., Nargang, F. E., Collins, R. A., and Zagozeski, C. A. (1977) Mol. Gen. Genet. 153, 247-257). The complementing wild-type gene has been cloned and sequenced, and shown to encode a protein with significant sequence similarity to Saccharomyces cerevisiae mitochondrial RNA polymerase and bacteriophage RNA polymerases. There are remarkable differences between the N. crassa protein and its yeast homologue, including a region of very little homology near the N termini of the two gene products. The cyt-5 gene encodes a stretch of polyglutamine in this region of unique sequence. In addition, an acidic insertion (86 amino acids, of which 24 are Asp or Glu and 10 are Arg or Lys) is present near the C terminus of the cyt-5 gene product. Transcript levels of the cytochrome b and cytochrome oxidase subunit III genes are severely reduced in cyt-5 mutants, suggesting a likely mechanism for the cytochrome-deficient phenotype. In contrast, mitochondrial rRNAs accumulate to nearly normal levels in cyt-5 mutants. However, mitochondrial rRNA levels are not indicative of the rate of transcription of the corresponding genes, since crude lysates of mitochondria from cyt-5 mutants exhibit greatly reduced transcriptional activity with a 19 S rRNA promoter. 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(Ohio State University, Columbus, OH.)</creatorcontrib><creatorcontrib>Kubelik, A.R</creatorcontrib><creatorcontrib>Mohr, S</creatorcontrib><creatorcontrib>Breitenberger, C.A</creatorcontrib><title>Cloning and characterization of the Neurospora crassa cyt-5 gene. A nuclear-coded mitochondrial RNA polymerase with a polyglutamine repeat</title><title>The Journal of biological chemistry</title><addtitle>J Biol Chem</addtitle><description>The Neurospora crassa mutants, cyt-5-1 and cyt-5-4, have a cytochrome b- and aa3-deficient phenotype, suggesting that they result from a deficiency in a nuclear-coded component of the mitochondrial gene expression apparatus (Bertrand, H., Nargang, F. E., Collins, R. A., and Zagozeski, C. A. (1977) Mol. Gen. Genet. 153, 247-257). The complementing wild-type gene has been cloned and sequenced, and shown to encode a protein with significant sequence similarity to Saccharomyces cerevisiae mitochondrial RNA polymerase and bacteriophage RNA polymerases. There are remarkable differences between the N. crassa protein and its yeast homologue, including a region of very little homology near the N termini of the two gene products. The cyt-5 gene encodes a stretch of polyglutamine in this region of unique sequence. In addition, an acidic insertion (86 amino acids, of which 24 are Asp or Glu and 10 are Arg or Lys) is present near the C terminus of the cyt-5 gene product. Transcript levels of the cytochrome b and cytochrome oxidase subunit III genes are severely reduced in cyt-5 mutants, suggesting a likely mechanism for the cytochrome-deficient phenotype. In contrast, mitochondrial rRNAs accumulate to nearly normal levels in cyt-5 mutants. However, mitochondrial rRNA levels are not indicative of the rate of transcription of the corresponding genes, since crude lysates of mitochondria from cyt-5 mutants exhibit greatly reduced transcriptional activity with a 19 S rRNA promoter. The cyt-5 gene is flanked by at least one gene whose product also may be involved in mitochondrial function</description><subject>Amino Acid Sequence</subject><subject>Cell Nucleus - enzymology</subject><subject>CITOCROMO B</subject><subject>CITOCROMOS</subject><subject>CLONACION</subject><subject>CLONAGE</subject><subject>Cloning, Molecular</subject><subject>COMPOSICION QUIMICA</subject><subject>COMPOSITION CHIMIQUE</subject><subject>CYTOCHROME</subject><subject>CYTOCHROME B</subject><subject>Cytochrome b Group - genetics</subject><subject>Cytochrome b Group - metabolism</subject><subject>DNA-Directed RNA Polymerases - genetics</subject><subject>Electron Transport Complex IV - genetics</subject><subject>Electron Transport Complex IV - metabolism</subject><subject>FENOTIPOS</subject><subject>GENE</subject><subject>Gene Expression</subject><subject>GENES</subject><subject>Genes, Fungal</subject><subject>Genetic Complementation Test</subject><subject>GENETICA</subject><subject>GENETIQUE</subject><subject>Mitochondria - enzymology</subject><subject>MITOCHONDRIE</subject><subject>MITOCONDRIA</subject><subject>Molecular Sequence Data</subject><subject>MUTANT</subject><subject>MUTANTES</subject><subject>Mutation</subject><subject>NEUROSPORA CRASSA</subject><subject>Neurospora crassa - enzymology</subject><subject>Neurospora crassa - genetics</subject><subject>Neurospora crassa - metabolism</subject><subject>Peptides - genetics</subject><subject>PHENOTYPE</subject><subject>Repetitive Sequences, Nucleic Acid</subject><subject>RNA, Fungal - genetics</subject><subject>RNA, Fungal - metabolism</subject><subject>Sequence Homology, Amino Acid</subject><subject>TRANSCRIPCION</subject><subject>TRANSCRIPTION</subject><subject>TRANSFERASAS</subject><subject>TRANSFERASE</subject><issn>0021-9258</issn><issn>1083-351X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkEFr3DAUhEVJSLZpz4VCQKfe7EiWJcvHZUnTQEghbaA381Z63lWwJUeSKZuf0F8d0-w97zLw5ps5DCFfOCs5a-qrp60pq4aXnJdKiuYDWXGmRSEk_3NCVoxVvGgrqc_Jx5Se2HJ1y8_ImVaVqqVekX-bIXjndxS8pWYPEUzG6F4gu-Bp6GneI73HOYY0hQjUREhpkUMuJN2hx5KuqZ_NgBALEyxaOroczD54Gx0M9OF-TacwHEZckkj_uryn8P-zG-YMo_NII04I-RM57WFI-PmoF-Tx-_XvzY_i7ufN7WZ9V_SVVLloa6u1UkyBFAyklcxoLcy2QmEEYz3o3vBGWr5ta221UUyAlLYH1SrBBBcX5Ntb7xTD84wpd6NLBocBPIY5dY1mnCvRvAvyhtWsreUCXh7BeTui7aboRoiH7rjy4n9983sIHeyiS93jr7bhtZateAVkSYhn</recordid><startdate>19960315</startdate><enddate>19960315</enddate><creator>Chen, B. 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(Ohio State University, Columbus, OH.) ; Kubelik, A.R ; Mohr, S ; Breitenberger, C.A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-f256t-94d886606a530a5d50c883cb2e3c300fa8fc175d1b948d8c603a55dfa69630313</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Amino Acid Sequence</topic><topic>Cell Nucleus - enzymology</topic><topic>CITOCROMO B</topic><topic>CITOCROMOS</topic><topic>CLONACION</topic><topic>CLONAGE</topic><topic>Cloning, Molecular</topic><topic>COMPOSICION QUIMICA</topic><topic>COMPOSITION CHIMIQUE</topic><topic>CYTOCHROME</topic><topic>CYTOCHROME B</topic><topic>Cytochrome b Group - genetics</topic><topic>Cytochrome b Group - metabolism</topic><topic>DNA-Directed RNA Polymerases - genetics</topic><topic>Electron Transport Complex IV - genetics</topic><topic>Electron Transport Complex IV - metabolism</topic><topic>FENOTIPOS</topic><topic>GENE</topic><topic>Gene Expression</topic><topic>GENES</topic><topic>Genes, Fungal</topic><topic>Genetic Complementation Test</topic><topic>GENETICA</topic><topic>GENETIQUE</topic><topic>Mitochondria - enzymology</topic><topic>MITOCHONDRIE</topic><topic>MITOCONDRIA</topic><topic>Molecular Sequence Data</topic><topic>MUTANT</topic><topic>MUTANTES</topic><topic>Mutation</topic><topic>NEUROSPORA CRASSA</topic><topic>Neurospora crassa - enzymology</topic><topic>Neurospora crassa - genetics</topic><topic>Neurospora crassa - metabolism</topic><topic>Peptides - genetics</topic><topic>PHENOTYPE</topic><topic>Repetitive Sequences, Nucleic Acid</topic><topic>RNA, Fungal - genetics</topic><topic>RNA, Fungal - metabolism</topic><topic>Sequence Homology, Amino Acid</topic><topic>TRANSCRIPCION</topic><topic>TRANSCRIPTION</topic><topic>TRANSFERASAS</topic><topic>TRANSFERASE</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chen, B. (Ohio State University, Columbus, OH.)</creatorcontrib><creatorcontrib>Kubelik, A.R</creatorcontrib><creatorcontrib>Mohr, S</creatorcontrib><creatorcontrib>Breitenberger, C.A</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Nucleic Acids 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>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The Journal of biological chemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chen, B. (Ohio State University, Columbus, OH.)</au><au>Kubelik, A.R</au><au>Mohr, S</au><au>Breitenberger, C.A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cloning and characterization of the Neurospora crassa cyt-5 gene. A nuclear-coded mitochondrial RNA polymerase with a polyglutamine repeat</atitle><jtitle>The Journal of biological chemistry</jtitle><addtitle>J Biol Chem</addtitle><date>1996-03-15</date><risdate>1996</risdate><volume>271</volume><issue>11</issue><spage>6537</spage><epage>6544</epage><pages>6537-6544</pages><issn>0021-9258</issn><eissn>1083-351X</eissn><abstract>The Neurospora crassa mutants, cyt-5-1 and cyt-5-4, have a cytochrome b- and aa3-deficient phenotype, suggesting that they result from a deficiency in a nuclear-coded component of the mitochondrial gene expression apparatus (Bertrand, H., Nargang, F. E., Collins, R. A., and Zagozeski, C. A. (1977) Mol. Gen. Genet. 153, 247-257). The complementing wild-type gene has been cloned and sequenced, and shown to encode a protein with significant sequence similarity to Saccharomyces cerevisiae mitochondrial RNA polymerase and bacteriophage RNA polymerases. There are remarkable differences between the N. crassa protein and its yeast homologue, including a region of very little homology near the N termini of the two gene products. The cyt-5 gene encodes a stretch of polyglutamine in this region of unique sequence. In addition, an acidic insertion (86 amino acids, of which 24 are Asp or Glu and 10 are Arg or Lys) is present near the C terminus of the cyt-5 gene product. Transcript levels of the cytochrome b and cytochrome oxidase subunit III genes are severely reduced in cyt-5 mutants, suggesting a likely mechanism for the cytochrome-deficient phenotype. In contrast, mitochondrial rRNAs accumulate to nearly normal levels in cyt-5 mutants. However, mitochondrial rRNA levels are not indicative of the rate of transcription of the corresponding genes, since crude lysates of mitochondria from cyt-5 mutants exhibit greatly reduced transcriptional activity with a 19 S rRNA promoter. The cyt-5 gene is flanked by at least one gene whose product also may be involved in mitochondrial function</abstract><cop>United States</cop><pmid>8626458</pmid><doi>10.1074/jbc.271.11.6537</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record>
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ispartof The Journal of biological chemistry, 1996-03, Vol.271 (11), p.6537-6544
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source MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection
subjects Amino Acid Sequence
Cell Nucleus - enzymology
CITOCROMO B
CITOCROMOS
CLONACION
CLONAGE
Cloning, Molecular
COMPOSICION QUIMICA
COMPOSITION CHIMIQUE
CYTOCHROME
CYTOCHROME B
Cytochrome b Group - genetics
Cytochrome b Group - metabolism
DNA-Directed RNA Polymerases - genetics
Electron Transport Complex IV - genetics
Electron Transport Complex IV - metabolism
FENOTIPOS
GENE
Gene Expression
GENES
Genes, Fungal
Genetic Complementation Test
GENETICA
GENETIQUE
Mitochondria - enzymology
MITOCHONDRIE
MITOCONDRIA
Molecular Sequence Data
MUTANT
MUTANTES
Mutation
NEUROSPORA CRASSA
Neurospora crassa - enzymology
Neurospora crassa - genetics
Neurospora crassa - metabolism
Peptides - genetics
PHENOTYPE
Repetitive Sequences, Nucleic Acid
RNA, Fungal - genetics
RNA, Fungal - metabolism
Sequence Homology, Amino Acid
TRANSCRIPCION
TRANSCRIPTION
TRANSFERASAS
TRANSFERASE
title Cloning and characterization of the Neurospora crassa cyt-5 gene. A nuclear-coded mitochondrial RNA polymerase with a polyglutamine repeat
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