Molecular cloning and identification of 3'-phosphoadenosine 5'-phosphosulfate transporter.
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.
Kamiyama
Shin
S
Division of Cell Biology, Soka University, 1-236 Tangi-cho, Hachioji, Tokyo 192-8577, Japan.
Suda
Takeshi
T
Ueda
Ryu
R
Suzuki
Masayuki
M
Okubo
Reiko
R
Kikuchi
Norihiro
N
Chiba
Yasunori
Y
Goto
Satoshi
S
Toyoda
Hidenao
H
Saigo
Kaoru
K
Watanabe
Masahiko
M
Narimatsu
Hisashi
H
Jigami
Yoshifumi
Y
Nishihara
Shoko
S
D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D003106:Colon / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D029721:Drosophila Proteins / Q000737:chemistry; D004331:Drosophila melanogaster; D006056:Golgi Apparatus / Q000378:metabolism; D006801:Humans; D026901:Membrane Transport Proteins / Q000737:chemistry* / Q000235:genetics*; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism; D010179:Pancreas / Q000378:metabolism; D010724:Phosphoadenosine Phosphosulfate / Q000378:metabolism*; D010802:Phylogeny; D010957:Plasmids / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013347:Subcellular Fractions; D013379:Substrate Specificity; D000076988:Sulfate Transporters; D014018:Tissue Distribution; D014162:Transfection
D018076:DNA, Complementary; D029721:Drosophila Proteins; D026901:Membrane Transport Proteins; D012330:RNA, Double-Stranded; C476248:SLC35B2 protein, human; D000076988:Sulfate Transporters; C476249:sll protein, Drosophila; D010724:Phosphoadenosine Phosphosulfate