title: Expression of the glucose transporter HXT1 involves the Ser-Thr protein phosphatase Sit4 in Saccharomyces cerevisiae.
abstract: We studied the effect of the loss of the Ser-Thr protein phosphatase Sit4, an important post-translational regulator, on the steady-state levels of the low-affinity glucose transporter Hxt1p and observed a delay in its appearance after high glucose induction, slow growth, and diminished glucose consumption. By analyzing the known essential pathway necessary to induce Hxt1p, we observed a partial inhibition of casein kinase I activity. In both WT and sit4Δ strains, the transcript was induced with no significant difference at 15 min of glucose induction; however, after 45 min, a clear difference in the level of expression was observed being 45% higher in WT than in sit4Δ strain. As at early time of induction, the HXT1 transcript was present but not the protein in the sit4Δ strain we analyzed association of HXT1 with ribosomes, which revealed a significant difference in the association profile; in the mutant strain, the HXT1 transcript associated with a larger set of ribosomal fractions than it did in the WT strain, suggesting also a partial defect in protein synthesis. Overexpression of the translation initiation factor TIF2/eIF4A led to an increase in Hxt1p abundance in the WT strain only. It was concluded that Sit4p ensures that HXT1 transcript is efficiently transcribed and translated thus increasing protein levels of Hxt1p when high glucose levels are present.
authors.affiliation: Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Brazil.
authors.forename: Andréa A
authors.initials: AA
authors.lastname: Souza
authors.forename: Michel N
authors.initials: MN
authors.lastname: Miranda
authors.forename: Suelene F
authors.initials: SF
authors.lastname: da Silva
authors.forename: Bruno
authors.initials: B
authors.lastname: Bozaquel-Morais
authors.forename: Claudio A
authors.initials: CA
authors.lastname: Masuda
authors.forename: Michel
authors.initials: M
authors.lastname: Ghislain
authors.forename: Mónica
authors.initials: M
authors.lastname: Montero-Lomelí
mesh_terms: D047389:Casein Kinase I / Q000378:metabolism; D039562:Eukaryotic Initiation Factor-4F / Q000235:genetics / Q000378:metabolism; D005285:Fermentation; D015966:Gene Expression Regulation, Fungal; D005947:Glucose / Q000378:metabolism; D051246:Glucose Transport Proteins, Facilitative / Q000235:genetics / Q000378:metabolism*; D015151:Immunoblotting; D010448:Peptide Initiation Factors / Q000235:genetics / Q000378:metabolism; D011132:Polyribosomes / Q000378:metabolism; D054648:Protein Phosphatase 2 / Q000235:genetics / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D013912:Threonine / Q000235:genetics / Q000378:metabolism
chemical_list: D039562:Eukaryotic Initiation Factor-4F; D051246:Glucose Transport Proteins, Facilitative; C090594:HXT1 protein, S cerevisiae; D010448:Peptide Initiation Factors; D029701:Saccharomyces cerevisiae Proteins; C504497:TIF2 protein, S cerevisiae; D013912:Threonine; D047389:Casein Kinase I; D054648:Protein Phosphatase 2; C104343:SIT4 protein, S cerevisiae; D005947:Glucose