Two Golgi-resident 3'-Phosphoadenosine 5'-phosphosulfate transporters play distinct roles in heparan sulfate modifications and embryonic and larval development in Caenorhabditis elegans.
Synthesis of extracellular sulfated molecules requires active 3'-phosphoadenosine 5'-phosphosulfate (PAPS). For sulfation to occur, PAPS must pass through the Golgi membrane, which is facilitated by Golgi-resident PAPS transporters. Caenorhabditis elegans PAPS transporters are encoded by two genes, pst-1 and pst-2. Using the yeast heterologous expression system, we characterized PST-1 and PST-2 as PAPS transporters. We created deletion mutants to study the importance of PAPS transporter activity. The pst-1 deletion mutant exhibited defects in cuticle formation, post-embryonic seam cell development, vulval morphogenesis, cell migration, and embryogenesis. The pst-2 mutant exhibited a wild-type phenotype. The defects observed in the pst-1 mutant could be rescued by transgenic expression of pst-1 and hPAPST1 but not pst-2 or hPAPST2. Moreover, the phenotype of a pst-1;pst-2 double mutant were similar to those of the pst-1 single mutant, except that larval cuticle formation was more severely defected. Disaccharide analysis revealed that heparan sulfate from these mutants was undersulfated. Gene expression reporter analysis revealed that these PAPS transporters exhibited different tissue distributions and subcellular localizations. These data suggest that pst-1 and pst-2 play different physiological roles in heparan sulfate modification and development.
Dejima
Katsufumi
K
Department of Biology, Faculty of Sciences 33, Kyushu University, Fukuoka 812-8581, Japan.
Murata
Daisuke
D
Mizuguchi
Souhei
S
Nomura
Kazuko H
KH
Izumikawa
Tomomi
T
Kitagawa
Hiroshi
H
Gengyo-Ando
Keiko
K
Yoshina
Sawako
S
Ichimiya
Tomomi
T
Nishihara
Shoko
S
Mitani
Shohei
S
Nomura
Kazuya
K
D000483:Alleles; D000818:Animals; D017173:Caenorhabditis elegans; D029742:Caenorhabditis elegans Proteins / Q000502:physiology*; D017353:Gene Deletion; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D006025:Glycosaminoglycans / Q000737:chemistry; D006056:Golgi Apparatus / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000737:chemistry; D006497:Heparan Sulfate / Q000378:metabolism*; D009154:Mutation; D033704:Nucleotide Transport Proteins / Q000502:physiology*; D013347:Subcellular Fractions; D013379:Substrate Specificity; D019076:Transgenes
D029742:Caenorhabditis elegans Proteins; D006025:Glycosaminoglycans; D049452:Green Fluorescent Proteins; D006497:Heparan Sulfate; D033704:Nucleotide Transport Proteins; C552207:Pst-1 protein, C elegans; C552208:Pst-2 protein, C elegans