title: Expression of the orphan cytosolic sulfotransferase SULT4A1 and its major splice variant in human tissues and cells: dimerization, degradation and polyubiquitination.
abstract: The cytosolic sulfotransferase SULT4A1 is highly conserved between mammalian species but its function remains unknown. Polymorphisms in the SULT4A1 gene have been linked to susceptibility to schizophrenia. There are 2 major SULT4A1 transcripts in humans, one that encodes full length protein (wild-type) and one that encodes a truncated protein (variant). Here, we investigated the expression of SULT4A1 in human tissues by RT-PCR and found the wild-type mRNA to be expressed mainly in the brain, gastrointestinal tract and prostate while the splice variant was more widely expressed. In human cell-lines, the wild-type transcript was found in neuronal cells, but the variant transcript was expressed in nearly all other lines examined. Western blot analysis only identified SULT4A1 protein in cells that expressed the wild-type mRNA. No variant protein was detected in cells that expressed the variant mRNA. Ectopically expressed full length SULT4A1 protein was stable while the truncated protein was not, having a half-life of approximately 3 hr. SULT4A1 was also shown to homodimerize, consistent with other SULTs that contain the consensus dimerization motif. Mutation of the dimerization motif resulted in a monomeric form of SULT4A1 that was rapidly degraded by polyubiquitination on the lysine located within the dimerization motif. These results show that SULT4A1 is widely expressed in human tissues, but mostly as a splice variant that produces a rapidly degraded protein. Dimerization protects the protein from degradation. Since many other cytosolic sulfotransferases possess the conserved lysine within the dimerization motif, homodimerization may serve, in part, to stabilize these enzymes in vivo.
authors.affiliation: Laboratory for Molecular and Cellular Pharmacology, School of Biomedical Sciences, University of Queensland, Brisbane, Australia.
authors.forename: Neelima P
authors.initials: NP
authors.lastname: Sidharthan
authors.affiliation: Laboratory for Molecular and Cellular Pharmacology, School of Biomedical Sciences, University of Queensland, Brisbane, Australia.
authors.forename: Neville J
authors.initials: NJ
authors.lastname: Butcher
authors.affiliation: Laboratory for Molecular and Cellular Pharmacology, School of Biomedical Sciences, University of Queensland, Brisbane, Australia.
authors.forename: Deanne J
authors.initials: DJ
authors.lastname: Mitchell
authors.affiliation: Laboratory for Molecular and Cellular Pharmacology, School of Biomedical Sciences, University of Queensland, Brisbane, Australia.
authors.forename: Rodney F
authors.initials: RF
authors.lastname: Minchin
mesh_terms: D000595:Amino Acid Sequence; D002460:Cell Line; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D025821:Polyubiquitin / Q000378:metabolism; D020033:Protein Isoforms / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D055503:Protein Multimerization; D055550:Protein Stability; D059748:Proteolysis; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D015238:Sulfotransferases / Q000032:analysis* / Q000235:genetics* / Q000378:metabolism; D054875:Ubiquitination
chemical_list: D020033:Protein Isoforms; D012333:RNA, Messenger; D025821:Polyubiquitin; C479953:SULT4A1 protein, human; D015238:Sulfotransferases