title: Mutant Potential Ubiquitination Sites in Dur3p Enhance the Urea and Ethyl Carbamate Reduction in a Model Rice Wine System.
abstract: Ubiquitination can significantly affect the endocytosis and degradation of plasma membrane proteins. Here, the ubiquitination of a Saccharomyces cerevisiae urea plasma membrane transporter (Dur3p) was altered. Two potential ubiquitination sites, lysine residues K556 and K571, of Dur3p were predicted and replaced by arginine, and the effects of these mutations on urea utilization and formation under different nitrogen conditions were investigated. Compared with Dur3p, the Dur3pK556Rmutant showed a 20.1% decrease in ubiquitination level in yeast nitrogen base medium containing urea and glutamine. It also exhibited a >75.8% decrease in urea formation in yeast extract-peptone-dextrose medium and 41.3 and 55.4% decreases in urea and ethyl carbamate formation (a known carcinogen), respectively, in a model rice wine system. The results presented here show that the mutation of Dur3p ubiquitination sites could significantly affect urea utilization and formation. Modifying the ubiquitination of specific transporters might have promising applications in rationally engineering S. cerevisiae strains to efficiently use specific nitrogen sources.
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University , 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University , 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
authors.forename: Guocheng
authors.initials: G
authors.lastname: Du
authors.affiliation: Zhejiang Guyuelongshan Shaoxing Wine Company , 13 Yangjiang Road, Shaoxing, Zhejiang 312099, China.
authors.forename: Huijun
authors.initials: H
authors.lastname: Zou
authors.affiliation: Zhejiang Guyuelongshan Shaoxing Wine Company , 13 Yangjiang Road, Shaoxing, Zhejiang 312099, China.
authors.forename: Guangfa
authors.initials: G
authors.lastname: Xie
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University , 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University , 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
authors.forename: Zhongping
authors.initials: Z
authors.lastname: Shi
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education and School of Biotechnology, Jiangnan University , 1800 Lihu Road, Wuxi, Jiangsu 214122, China.
authors.forename: Jingwen
authors.identifier: http://orcid.org/0000-0002-3949-3733
authors.initials: J
authors.lastname: Zhou
mesh_terms: D020816:Amino Acid Motifs; D005285:Fermentation; D005973:Glutamine / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012275:Oryza / Q000378:metabolism / Q000382:microbiology*; D010084:Oxidation-Reduction; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D054875:Ubiquitination; D014508:Urea / Q000378:metabolism*; D014520:Urethane / Q000378:metabolism*; D014920:Wine / Q000032:analysis / Q000382:microbiology*
chemical_list: C515373:DUR3 protein, S cerevisiae; D026901:Membrane Transport Proteins; D029701:Saccharomyces cerevisiae Proteins; D005973:Glutamine; D014520:Urethane; D014508:Urea
keywords: nitrogen catabolite repression; nitrogen sources; post-translational modifications; site-directed mutagenesis; urea transporter