Cloning and expression of a yeast ubiquitin-protein cleaving activity in Escherichia coli
We have cloned the gene coding for a ubiquitin-protein hydrolase from the yeast Saccharomyces cerevisiae . The gene ( YUH1 ) was isolated from a yeast genomic library by screening with an oligonucleotide probe designed from the amino acid sequence of the purified hydrolase. The YUH1 gene encodes a 2...
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| Veröffentlicht in: | Bio/Technology 1989-07, Vol.7 (7), p.698-704 |
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| creator | Miller, H.I. (Gentech Inc., South San Francisco, CA.) Henzel, W.J Ridgway, J.B Kuang, W.J Chisholm, V Liu, C.C |
| description | We have cloned the gene coding for a ubiquitin-protein hydrolase from the yeast
Saccharomyces cerevisiae
. The gene (
YUH1
) was isolated from a yeast genomic library by screening with an oligonucleotide probe designed from the amino acid sequence of the purified hydrolase. The
YUH1
gene encodes a 26 kD protein and contains no introns. The
YUH1
gene product can be overexpressed in active form in
Escherichia coli
and purified by a two column procedure. The purified hydrolase is capable of cleaving ubiquitin-protein fusions
in vitro
specifically at the ubiquitin fusion junction and requires no high energy cofactors. Fusions can also be cleaved in
E. coli
in strains expressing the hydrolase. Gene disruptions in haploid yeast strains have no apparent phenotypic change and ubiquitin-protein hydrolase activity in extracts is normal, indicating the existence of additional genes for ubiquitin-protein hydrolase.
In vitro
and
in vivo
cleavage of ubiquitin-protein fusions may be a useful method of producing proteins with defined ammo-termini. |
| doi_str_mv | 10.1038/nbt0789-698 |
| format | Article |
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Saccharomyces cerevisiae
. The gene (
YUH1
) was isolated from a yeast genomic library by screening with an oligonucleotide probe designed from the amino acid sequence of the purified hydrolase. The
YUH1
gene encodes a 26 kD protein and contains no introns. The
YUH1
gene product can be overexpressed in active form in
Escherichia coli
and purified by a two column procedure. The purified hydrolase is capable of cleaving ubiquitin-protein fusions
in vitro
specifically at the ubiquitin fusion junction and requires no high energy cofactors. Fusions can also be cleaved in
E. coli
in strains expressing the hydrolase. Gene disruptions in haploid yeast strains have no apparent phenotypic change and ubiquitin-protein hydrolase activity in extracts is normal, indicating the existence of additional genes for ubiquitin-protein hydrolase.
In vitro
and
in vivo
cleavage of ubiquitin-protein fusions may be a useful method of producing proteins with defined ammo-termini.</description><identifier>ISSN: 0733-222X</identifier><identifier>ISSN: 1087-0156</identifier><identifier>EISSN: 2331-3684</identifier><identifier>EISSN: 1546-1696</identifier><identifier>DOI: 10.1038/nbt0789-698</identifier><language>eng</language><publisher>New York: Nature Publishing Group US</publisher><subject>Agriculture ; AMINO ACID SEQUENCES ; Bioinformatics ; Biological and medical sciences ; Biomedical and Life Sciences ; Biomedical Engineering/Biotechnology ; Biomedicine ; Biotechnology ; CLONACION ; CLONAGE ; CLONING ; ESCHERICHIA COLI ; EXPRESION GENICA ; EXPRESSION DES GENES ; Fundamental and applied biological sciences. Psychology ; GENE ; GENE EXPRESSION ; GENE TRANSFER ; GENES ; Genetic engineering ; Genetic technics ; GENETIC TRANSFORMATION ; HYDRO-LYASES ; LIASAS ; Life Sciences ; LYASE ; LYASES ; Methods. Procedures. Technologies ; Molecular cloning ; MOLECULAR SEQUENCE DATA ; NUCLEOTIDE SEQUENCE ; research-paper ; SACCHAROMYCES CEREVISIAE ; SECUENCIA NUCLEICA ; SEQUENCE NUCLEIQUE ; STRUCTURAL GENES ; TRANSFERENCIA DE GENES ; TRANSFERT DE GENE ; TRANSFORMACION GENETICA ; TRANSFORMATION GENETIQUE ; YUH1 GENE</subject><ispartof>Bio/Technology, 1989-07, Vol.7 (7), p.698-704</ispartof><rights>Nature Publishing Company 1989</rights><rights>1989 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c352t-64185b17e6e8c73a9b8895f2df55f18afd0a1c75367b0c2605a1119f9d735b3f3</citedby><cites>FETCH-LOGICAL-c352t-64185b17e6e8c73a9b8895f2df55f18afd0a1c75367b0c2605a1119f9d735b3f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/nbt0789-698$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/nbt0789-698$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,777,781,27905,27906,41469,42538,51300</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=7344972$$DView record in Pascal Francis$$Hfree_for_read</backlink></links><search><creatorcontrib>Miller, H.I. (Gentech Inc., South San Francisco, CA.)</creatorcontrib><creatorcontrib>Henzel, W.J</creatorcontrib><creatorcontrib>Ridgway, J.B</creatorcontrib><creatorcontrib>Kuang, W.J</creatorcontrib><creatorcontrib>Chisholm, V</creatorcontrib><creatorcontrib>Liu, C.C</creatorcontrib><title>Cloning and expression of a yeast ubiquitin-protein cleaving activity in Escherichia coli</title><title>Bio/Technology</title><addtitle>Nat Biotechnol</addtitle><description>We have cloned the gene coding for a ubiquitin-protein hydrolase from the yeast
Saccharomyces cerevisiae
. The gene (
YUH1
) was isolated from a yeast genomic library by screening with an oligonucleotide probe designed from the amino acid sequence of the purified hydrolase. The
YUH1
gene encodes a 26 kD protein and contains no introns. The
YUH1
gene product can be overexpressed in active form in
Escherichia coli
and purified by a two column procedure. The purified hydrolase is capable of cleaving ubiquitin-protein fusions
in vitro
specifically at the ubiquitin fusion junction and requires no high energy cofactors. Fusions can also be cleaved in
E. coli
in strains expressing the hydrolase. Gene disruptions in haploid yeast strains have no apparent phenotypic change and ubiquitin-protein hydrolase activity in extracts is normal, indicating the existence of additional genes for ubiquitin-protein hydrolase.
In vitro
and
in vivo
cleavage of ubiquitin-protein fusions may be a useful method of producing proteins with defined ammo-termini.</description><subject>Agriculture</subject><subject>AMINO ACID SEQUENCES</subject><subject>Bioinformatics</subject><subject>Biological and medical sciences</subject><subject>Biomedical and Life Sciences</subject><subject>Biomedical Engineering/Biotechnology</subject><subject>Biomedicine</subject><subject>Biotechnology</subject><subject>CLONACION</subject><subject>CLONAGE</subject><subject>CLONING</subject><subject>ESCHERICHIA COLI</subject><subject>EXPRESION GENICA</subject><subject>EXPRESSION DES GENES</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>GENE</subject><subject>GENE EXPRESSION</subject><subject>GENE TRANSFER</subject><subject>GENES</subject><subject>Genetic engineering</subject><subject>Genetic technics</subject><subject>GENETIC TRANSFORMATION</subject><subject>HYDRO-LYASES</subject><subject>LIASAS</subject><subject>Life Sciences</subject><subject>LYASE</subject><subject>LYASES</subject><subject>Methods. Procedures. Technologies</subject><subject>Molecular cloning</subject><subject>MOLECULAR SEQUENCE DATA</subject><subject>NUCLEOTIDE SEQUENCE</subject><subject>research-paper</subject><subject>SACCHAROMYCES CEREVISIAE</subject><subject>SECUENCIA NUCLEICA</subject><subject>SEQUENCE NUCLEIQUE</subject><subject>STRUCTURAL GENES</subject><subject>TRANSFERENCIA DE GENES</subject><subject>TRANSFERT DE GENE</subject><subject>TRANSFORMACION GENETICA</subject><subject>TRANSFORMATION GENETIQUE</subject><subject>YUH1 GENE</subject><issn>0733-222X</issn><issn>1087-0156</issn><issn>2331-3684</issn><issn>1546-1696</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1989</creationdate><recordtype>article</recordtype><recordid>eNp1kD1PwzAQhi0EEqUwsTF5YIOAP-LYHlFVPqRKDFAJpuji2K2r4BQ7rei_J5CKjemku-d9pXsQOqfkhhKubkPVEal0Vmh1gEaMc5rxQuWHaEQk5xlj7O0YnaS0IiSXBctH6H3StMGHBYZQY_u1jjYl3wbcOgx4ZyF1eFP5z43vfMjWse2sD9g0Fra_IdP5re92uF9Ok1na6M3SAzZt40_RkYMm2bP9HKP5_fR18pjNnh-eJnezzHDBuqzIqRIVlbawykgOulJKC8dqJ4SjClxNgBopeCErYlhBBFBKtdO15KLijo_R1dBrYptStK5cR_8BcVdSUv5YKfdWyt5KT18O9BqSgcZFCManv4jkea4l67HrAUv9JSxsLFftJob-j39aLwbcQVvCIvaN8xctmBK54N-mynn4</recordid><startdate>19890701</startdate><enddate>19890701</enddate><creator>Miller, H.I. (Gentech Inc., South San Francisco, CA.)</creator><creator>Henzel, W.J</creator><creator>Ridgway, J.B</creator><creator>Kuang, W.J</creator><creator>Chisholm, V</creator><creator>Liu, C.C</creator><general>Nature Publishing Group US</general><general>Nature Publications</general><scope>FBQ</scope><scope>IQODW</scope><scope>AAYXX</scope><scope>CITATION</scope></search><sort><creationdate>19890701</creationdate><title>Cloning and expression of a yeast ubiquitin-protein cleaving activity in Escherichia coli</title><author>Miller, H.I. (Gentech Inc., South San Francisco, CA.) ; Henzel, W.J ; Ridgway, J.B ; Kuang, W.J ; Chisholm, V ; Liu, C.C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c352t-64185b17e6e8c73a9b8895f2df55f18afd0a1c75367b0c2605a1119f9d735b3f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1989</creationdate><topic>Agriculture</topic><topic>AMINO ACID SEQUENCES</topic><topic>Bioinformatics</topic><topic>Biological and medical sciences</topic><topic>Biomedical and Life Sciences</topic><topic>Biomedical Engineering/Biotechnology</topic><topic>Biomedicine</topic><topic>Biotechnology</topic><topic>CLONACION</topic><topic>CLONAGE</topic><topic>CLONING</topic><topic>ESCHERICHIA COLI</topic><topic>EXPRESION GENICA</topic><topic>EXPRESSION DES GENES</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>GENE</topic><topic>GENE EXPRESSION</topic><topic>GENE TRANSFER</topic><topic>GENES</topic><topic>Genetic engineering</topic><topic>Genetic technics</topic><topic>GENETIC TRANSFORMATION</topic><topic>HYDRO-LYASES</topic><topic>LIASAS</topic><topic>Life Sciences</topic><topic>LYASE</topic><topic>LYASES</topic><topic>Methods. Procedures. Technologies</topic><topic>Molecular cloning</topic><topic>MOLECULAR SEQUENCE DATA</topic><topic>NUCLEOTIDE SEQUENCE</topic><topic>research-paper</topic><topic>SACCHAROMYCES CEREVISIAE</topic><topic>SECUENCIA NUCLEICA</topic><topic>SEQUENCE NUCLEIQUE</topic><topic>STRUCTURAL GENES</topic><topic>TRANSFERENCIA DE GENES</topic><topic>TRANSFERT DE GENE</topic><topic>TRANSFORMACION GENETICA</topic><topic>TRANSFORMATION GENETIQUE</topic><topic>YUH1 GENE</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Miller, H.I. (Gentech Inc., South San Francisco, CA.)</creatorcontrib><creatorcontrib>Henzel, W.J</creatorcontrib><creatorcontrib>Ridgway, J.B</creatorcontrib><creatorcontrib>Kuang, W.J</creatorcontrib><creatorcontrib>Chisholm, V</creatorcontrib><creatorcontrib>Liu, C.C</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>CrossRef</collection><jtitle>Bio/Technology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Miller, H.I. (Gentech Inc., South San Francisco, CA.)</au><au>Henzel, W.J</au><au>Ridgway, J.B</au><au>Kuang, W.J</au><au>Chisholm, V</au><au>Liu, C.C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cloning and expression of a yeast ubiquitin-protein cleaving activity in Escherichia coli</atitle><jtitle>Bio/Technology</jtitle><stitle>Nat Biotechnol</stitle><date>1989-07-01</date><risdate>1989</risdate><volume>7</volume><issue>7</issue><spage>698</spage><epage>704</epage><pages>698-704</pages><issn>0733-222X</issn><issn>1087-0156</issn><eissn>2331-3684</eissn><eissn>1546-1696</eissn><abstract>We have cloned the gene coding for a ubiquitin-protein hydrolase from the yeast
Saccharomyces cerevisiae
. The gene (
YUH1
) was isolated from a yeast genomic library by screening with an oligonucleotide probe designed from the amino acid sequence of the purified hydrolase. The
YUH1
gene encodes a 26 kD protein and contains no introns. The
YUH1
gene product can be overexpressed in active form in
Escherichia coli
and purified by a two column procedure. The purified hydrolase is capable of cleaving ubiquitin-protein fusions
in vitro
specifically at the ubiquitin fusion junction and requires no high energy cofactors. Fusions can also be cleaved in
E. coli
in strains expressing the hydrolase. Gene disruptions in haploid yeast strains have no apparent phenotypic change and ubiquitin-protein hydrolase activity in extracts is normal, indicating the existence of additional genes for ubiquitin-protein hydrolase.
In vitro
and
in vivo
cleavage of ubiquitin-protein fusions may be a useful method of producing proteins with defined ammo-termini.</abstract><cop>New York</cop><pub>Nature Publishing Group US</pub><doi>10.1038/nbt0789-698</doi><tpages>7</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0733-222X |
| ispartof | Bio/Technology, 1989-07, Vol.7 (7), p.698-704 |
| issn | 0733-222X 1087-0156 2331-3684 1546-1696 |
| language | eng |
| recordid | cdi_crossref_primary_10_1038_nbt0789_698 |
| source | Springer Nature - Complete Springer Journals; Nature Journals Online |
| subjects | Agriculture AMINO ACID SEQUENCES Bioinformatics Biological and medical sciences Biomedical and Life Sciences Biomedical Engineering/Biotechnology Biomedicine Biotechnology CLONACION CLONAGE CLONING ESCHERICHIA COLI EXPRESION GENICA EXPRESSION DES GENES Fundamental and applied biological sciences. Psychology GENE GENE EXPRESSION GENE TRANSFER GENES Genetic engineering Genetic technics GENETIC TRANSFORMATION HYDRO-LYASES LIASAS Life Sciences LYASE LYASES Methods. Procedures. Technologies Molecular cloning MOLECULAR SEQUENCE DATA NUCLEOTIDE SEQUENCE research-paper SACCHAROMYCES CEREVISIAE SECUENCIA NUCLEICA SEQUENCE NUCLEIQUE STRUCTURAL GENES TRANSFERENCIA DE GENES TRANSFERT DE GENE TRANSFORMACION GENETICA TRANSFORMATION GENETIQUE YUH1 GENE |
| title | Cloning and expression of a yeast ubiquitin-protein cleaving activity in Escherichia coli |
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