title: Yeast Actin-Related Protein ARP6 Negatively Regulates Agrobacterium-Mediated Transformation of Yeast Cell.
abstract: The yeasts, including Saccharomyces cerevisiae and Pichia pastoris, are single-cell eukaryotic organisms that can serve as models for human genetic diseases and hosts for large scale production of recombinant proteins in current biopharmaceutical industry. Thus, efficient genetic engineering tools for yeasts are of great research and economic values. Agrobacterium tumefaciens-mediated transformation (AMT) can transfer T-DNA into yeast cells as a method for genetic engineering. However, how the T-DNA is transferred into the yeast cells is not well established yet. Here our genetic screening of yeast knockout mutants identified a yeast actin-related protein ARP6 as a negative regulator of AMT. ARP6 is a critical member of the SWR1 chromatin remodeling complex (SWR-C); knocking out some other components of the complex also increased the transformation efficiency, suggesting that ARP6 might regulate AMT via SWR-C. Moreover, knockout of ARP6 led to disruption of microtubule integrity, higher uptake and degradation of virulence proteins, and increased DNA stability inside the cells, all of which resulted in enhanced transformation efficiency. Our findings have identified molecular and cellular mechanisms regulating AMT and a potential target for enhancing the transformation efficiency in yeast cells.
authors.affiliation: Key Laboratory for Major Obstetric Diseases of Guangdong Province, Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, China.
authors.forename: Yumei
authors.initials: Y
authors.lastname: Luo
authors.affiliation: Department of Biological Sciences, National University of Singapore, Singapore 117543 ; Faculty of Health Sciences, University of Macau, Macau.
authors.forename: Zikai
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Department of Biological Sciences, National University of Singapore, Singapore 117543 ; Faculty of Health Sciences, University of Macau, Macau.
authors.forename: Detu
authors.initials: D
authors.lastname: Zhu
authors.affiliation: Department of Biological Sciences, National University of Singapore, Singapore 117543.
authors.forename: Haitao
authors.initials: H
authors.lastname: Tu
authors.affiliation: Department of Biological Sciences, National University of Singapore, Singapore 117543.
authors.forename: Shen Quan
authors.initials: SQ
authors.lastname: Pan
mesh_terms: D000199:Actins / Q000378:metabolism*; D016960:Agrobacterium tumefaciens / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D042002:Chromatin Assembly and Disassembly / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism; D003851:Deoxyribonucleases / Q000378:metabolism; D055786:Gene Knockout Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007473:Ion Channels / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D059748:Proteolysis; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D014170:Transformation, Genetic
chemical_list: C000609084:ARP6 protein, S cerevisiae; D000199:Actins; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D007473:Ion Channels; D029701:Saccharomyces cerevisiae Proteins; C036483:T-DNA; C051583:virE2 protein, Agrobacterium; D049452:Green Fluorescent Proteins; D003851:Deoxyribonucleases