Molecular cloning and identification of 3'-phosphoadenosine 5'-phosphosulfate transporter

J Biol Chem. 2003 Jul 11;278(28):25958-63. doi: 10.1074/jbc.M302439200. Epub 2003 Apr 25.

Abstract

Nucleotide sulfate, namely 3'-phosphoadenosine 5'-phosphosulfate (PAPS), is a universal sulfuryl donor for sulfation. Although a specific PAPS transporter is present in Golgi membrane, no study has reported the corresponding gene. We have identified a novel human gene encoding a PAPS transporter, which we have named PAPST1, and the Drosophila melanogaster ortholog, slalom (sll). The amino acid sequence of PAPST1 (432 amino acids) exhibited 48.1% identity with SLL (465 amino acids), and hydropathy analysis predicted the two to be type III transmembrane proteins. The transient expression of PAPST1 in SW480 cells showed a subcellular localization in Golgi membrane. The expression of PAPST1 and SLL in yeast Saccharomyces cerevisiae significantly increased the transport of PAPS into the Golgi membrane fraction. In human tissues, PAPST1 is highly expressed in the placenta and pancreas and present at lower levels in the colon and heart. An RNA interference fly of sll produced with a GAL4-UAS system revealed that the PAPS transporter is essential for viability. It is well known that mutations of some genes related to PAPS synthesis are responsible for human inherited disorders. Our findings provide insights into the significance of PAPS transport and post-translational sulfation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Biological Transport
  • Blotting, Western
  • Cell Membrane / metabolism
  • Cloning, Molecular
  • Colon / metabolism
  • DNA, Complementary / metabolism
  • Dose-Response Relationship, Drug
  • Drosophila Proteins / chemistry
  • Drosophila melanogaster
  • Golgi Apparatus / metabolism
  • Humans
  • Membrane Transport Proteins / chemistry*
  • Membrane Transport Proteins / genetics*
  • Microscopy, Fluorescence
  • Molecular Sequence Data
  • Myocardium / metabolism
  • Pancreas / metabolism
  • Phosphoadenosine Phosphosulfate / metabolism*
  • Phylogeny
  • Plasmids / metabolism
  • Protein Processing, Post-Translational
  • RNA Interference
  • RNA, Double-Stranded / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Saccharomyces cerevisiae / metabolism
  • Sequence Homology, Amino Acid
  • Subcellular Fractions
  • Substrate Specificity
  • Sulfate Transporters
  • Tissue Distribution
  • Transfection

Substances

  • DNA, Complementary
  • Drosophila Proteins
  • Membrane Transport Proteins
  • RNA, Double-Stranded
  • SLC35B2 protein, human
  • Sulfate Transporters
  • sll protein, Drosophila
  • Phosphoadenosine Phosphosulfate

Associated data

  • GENBANK/AB106538
  • GENBANK/AB107958