title: Differential expression of a C-terminal splice variant of phosphatidylinositol transfer protein beta lacking the constitutive-phosphorylated Ser262 that localizes to the Golgi compartment.
abstract: Mammalian PITPbeta (phosphatidylinositol transfer protein beta) is a 272-amino-acid polypeptide capable of transferring PtdIns, PtdCho and SM (sphingomyelin) between membrane bilayers. It has been reported that Ser262 present in the C-terminus of PITPbeta is constitutively phosphorylated and determines Golgi localization. We provide evidence for the expression of an sp (splice) variant of PITPbeta (PITPbeta-sp2) where the C-terminal 15 amino acids of PITPbeta-sp1 are replaced by an alternative C-terminus of 16 amino acids. PITPbeta-sp1 is the product of the first 11 exons, whereas PITPbeta-sp2 is a product of the first 10 exons followed by the twelfth exon--exon 11 being 'skipped'. Both splice variants are capable of PtdIns and PtdCho transfer, with PITPbeta-sp2 being unable to transport SM. PITPbeta is ubiquitously expressed, with the highest amounts of PITPbeta found in HL60 cells and in rat liver; HL60 cells express only PITPbeta-sp1, whereas rat liver expresses both sp variants in similar amounts. In both cell types, PITPbeta-sp1 is constitutively phosphorylated and both the PtdIns and PtdCho forms of PITPbeta-sp1 are present. In contrast, PITPbeta-sp2 lacks the constitutively phosphorylated Ser262 (replaced with glutamine). Nonetheless, both PITPbeta variants localize to the Golgi and, moreover, dephosphorylation of Ser262 of PITPbeta-sp1 does not affect its Golgi localization. The presence of PITPbeta sp variants adds an extra level of proteome complexity and, in rat liver, the single gene for PITPbeta gives rise to seven distinct protein species that can be resolved on the basis of their charge differences.
authors.affiliation: Lipid Signalling Group, Department of Physiology, University College London, 21 University Street, Rockefeller Building, London WC1E 6JJ, UK.
authors.forename: Clive P
authors.initials: CP
authors.lastname: Morgan
authors.forename: Victoria
authors.initials: V
authors.lastname: Allen-Baume
authors.forename: Marko
authors.initials: M
authors.lastname: Radulovic
authors.forename: Michelle
authors.initials: M
authors.lastname: Li
authors.forename: Alison
authors.initials: A
authors.lastname: Skippen
authors.forename: Shamshad
authors.initials: S
authors.lastname: Cockcroft
mesh_terms: D017398:Alternative Splicing / Q000502:physiology*; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D003600:Cytosol; D005786:Gene Expression Regulation; D006056:Golgi Apparatus / Q000378:metabolism*; D018922:HL-60 Cells; D006801:Humans; D008099:Liver / Q000166:cytology; D008969:Molecular Sequence Data; D049294:Phospholipid Transfer Proteins / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D021381:Protein Transport; D051381:Rats
chemical_list: D049294:Phospholipid Transfer Proteins