title: Expression of FLR1 transporter requires phospholipase C and is repressed by Mediator.
abstract: In budding yeast, phosphoinositide-specific phospholipase C (Plc1p encoded by PLC1 gene) is important for function of kinetochores. Deletion of PLC1 results in benomyl sensitivity, alterations in chromatin structure of centromeres, mitotic delay, and a higher frequency of chromosome loss. Here we intended to utilize benomyl sensitivity as a phenotype that would allow us to identify genes that are important for kinetochore function and are downstream of Plc1p. However, our screen identified SIN4, encoding a component of the Mediator complex of RNA polymerase II. Deletion of SIN4 gene (sin4Delta) does not suppress benomyl sensitivity of plc1Delta cells by improving the function of kinetochores. Instead, benomyl sensitivity of plc1Delta cells is caused by a defect in expression of FLR1, and the suppression of benomyl sensitivity in plc1Delta sin4Delta cells occurs by derepression of FLR1 transcription. FLR1 encodes a plasma membrane transporter that mediates resistance to benomyl. Several other mutations in the Mediator complex also result in significant derepression of FLR1 and greatly increased resistance to benomyl. Thus, benomyl sensitivity is not a phenotype exclusively associated with mitotic spindle defect. These results demonstrate that in addition to promoter-specific transcription factors that are components of the pleiotropic drug resistance network, expression of the membrane transporters can be regulated by Plc1p, a component of a signal transduction pathway, and by Mediator, a general transcription factor. The results thus suggest another layer of complexity in regulation of pleiotropic drug resistance.
authors.affiliation: Department of Biological Sciences, St. John's University, 8000 Utopia Parkway, Queens, NY 11439, USA.
authors.forename: Carlos
authors.initials: C
authors.lastname: Romero
authors.forename: Parima
authors.initials: P
authors.lastname: Desai
authors.forename: Nicholas
authors.initials: N
authors.lastname: DeLillo
authors.forename: Ales
authors.initials: A
authors.lastname: Vancura
mesh_terms: D001542:Benomyl / Q000494:pharmacology; D025141:Drug Resistance, Fungal / Q000502:physiology; D005659:Fungicides, Industrial / Q000494:pharmacology; D015966:Gene Expression Regulation, Fungal; D018386:Kinetochores / Q000378:metabolism; D056892:Mediator Complex; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D046912:Multiprotein Complexes; D027361:Organic Anion Transporters; D010641:Phenotype; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D010738:Type C Phospholipases / Q000378:metabolism*
chemical_list: C118429:FLR1 protein, S cerevisiae; D005659:Fungicides, Industrial; D056892:Mediator Complex; D026901:Membrane Transport Proteins; D046912:Multiprotein Complexes; D027361:Organic Anion Transporters; C076878:SIN4 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D015534:Trans-Activators; D010738:Type C Phospholipases; D001542:Benomyl