Proteoglycan UDP-Galactose:β-Xylose β1,4-Galactosyltransferase I Is Essential for Viability inDrosophila melanogaster

Takemae, Hitoshi; Ueda, Ryo; Okubo, Reiko; Nakato, Hiroshi; Izumi, Susumu; Saigo, Kaoru; Nishihara, Shoko

doi:10.1074/jbc.m301123200

Cited by 42 publications

(26 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Indeed, sll in the flybase is involved in signal transduction by growth factors of Wingless and Hedgehog during patterning and morphogenesis (39). We reported the requirement of Drosophila ␤1,4-galactosyltransferase I, which contributes to the synthesis of the linkage structure of proteoglycans, for viability (21). However, we did not investigate whether the lethality of the sll RNAi fly is caused by a reduction in the sulfation of proteoglycans or other sulfated glycoconjugates, such as sulfatides (40) and HNK-1 epitope (41).…”

Section: Fig 2 Subcellular Localization Of Papst1 Proteinmentioning

confidence: 94%

See 1 more Smart Citation

Molecular Cloning and Identification of 3′-Phosphoadenosine 5′-Phosphosulfate Transporter

Kamiyama

Suda

Ueda

et al. 2003

Journal of Biological Chemistry

Self Cite

127

View full text Add to dashboard Cite

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.Glycosylation, phosphorylation, and sulfation are essential post-translational alterations of glycoproteins, proteoglycans, and glycolipids for normal growth and development. For sulfation, an activated form of sulfate, 3Ј-phosphoadenosine 5Ј-phosphosulfate (PAPS), 1 is used as a common sulfate donor (1). Sulfate is transferred from PAPS to a defined position on the sugar residue by sulfotransferases. In higher organisms, PAPS is synthesized in the cytosol (2, 3) by a bifunctional PAPS synthetase having both ATP-sulfurylase and adenosine-phosphosulfate kinase activities (4). Since sulfation occurs in the lumens of the endoplasmic reticulum and Golgi apparatus (5), PAPS must be translocated from the cytosol into the Golgi lumen through a specific transporter localized in the microsomal membrane.Earlier studies had shown a saturable transport activity of PAPS using isolated Golgi vesicles (6) or reconstituted proteoliposome (7). To identify this transporter protein, the proteins responsible for PAPS translocating activity have been purified (8 -10). The characterization of these purified proteins revealed the kinetic behavior of a PAPS-specific transport through an antiport mechanism (8 -10); however, cloning of the transporter has not been reported.Recently, several nucleotide-sugar transporters (NSTs) have been cloned and characterized in mammals, yeast, protozoa, and plants (11-13). Nucleotide-sugars are the donor substrates for glycosylation, which is catalyzed by glycosyltransferases. These NST proteins are highly hydrophobic Type III multitransmembrane proteins localized in the Golgi or endoplasmic reticulum membrane and provide a specific substrate for the glycosylation. The structural conservation among NSTs has contributed to the identifi...

show abstract

Section: Fig 2 Subcellular Localization Of Papst1 Proteinmentioning

confidence: 94%

“…The cloning of sll into the transformation vector pUAST was done as previously reported (21). The transformation of Drosophila embryos was carried out according to Spradling (22) with w 1118 stock as a host to make four UAS-sll inverted repeat fly lines.…”

Section: Isolation Of Human and Drosophila Paps Transporter Cdnas Andmentioning

confidence: 99%

Molecular Cloning and Identification of 3′-Phosphoadenosine 5′-Phosphosulfate Transporter

Kamiyama

Suda

Ueda

et al. 2003

Journal of Biological Chemistry

Self Cite

127

View full text Add to dashboard Cite

show abstract

“…RNAi knockdown was performed with a heritable and inducible RNAi system using the GAL4-UAS-IR system (23,24). The mRNA levels of dPOMT1 and dPOMT2 in each of the RNAi flies were reduced to 32 and 21% of those in wild-type flies, respectively, and no effects of RNAi on the mRNA levels of the other members of the dPOMT family were observed (Fig.…”

Section: Discussionmentioning

confidence: 99%

The Twisted Abdomen Phenotype of Drosophila POMT1 and POMT2 Mutants Coincides with Their Heterophilic Protein O-Mannosyltransferase Activity

Ichimiya

Manya

Ohmae

et al. 2004

Journal of Biological Chemistry

Self Cite

110

View full text Add to dashboard Cite

Walker-Warburg syndrome, caused by mutations in protein O-mannosyltransferase-1 (POMT1), is an autosomal recessive disorder characterized by severe brain malformation, muscular dystrophy, and structural eye abnormalities. As humans have a second POMT, POMT2, we cloned each Drosophila ortholog of the human POMT genes and carried out RNA interference (RNAi) knockdown to investigate the function of these proteins in vivo. Drosophila POMT2 (dPOMT2) RNAi mutant flies showed a "twisted abdomen phenotype," in which the abdomen is twisted 30 -60°, similar to the dPOMT1 mutant. Moreover, dPOMT2 interacted genetically with dPOMT1, suggesting that the dPOMTs function in collaboration with each other in vivo. We expressed dPOMTs in Sf21 cells and measured POMT activity. dPOMT2 transferred a mannose to the dystroglycan protein only when it was coexpressed with dPOMT1. Likewise, dPOMT1 showed POMT activity only when coexpressed with dPOMT2, and neither dPOMT showed any activity by itself. Each dPOMT RNAi fly totally reduced POMT activity, despite the specific reduction in the level of each dPOMT mRNA. The expression pattern of dPOMT2 mRNA was found to be similar to that of dPOMT1 mRNA using whole mount in situ hybridization. These results demonstrate that the two dPOMTs function as a protein O-mannosyltransferase in association with each other, in vitro and in vivo, to generate and maintain normal muscle development.

show abstract

“…2A) (10)(11)(12). HS has repeating disaccharide units of D-glucuronic acid-N-acetyl-D-glucosamine (GlcA-GlcNAc) that are modiˆed diŠerentially by epimerization and sulfation.…”

Section: Heparan Sulfate (Hs) Is Expressed On Es Cellsmentioning

confidence: 99%

“…The eŠect of HS on self-renewal and proliferation in mouse ES cells. 2A) (10)(11)(12). We therefore knocked-down expression of EXT1 using RNA interference (RNAi) to determine the eŠect of a reduction in HS production on mouse ES cells, with particular attention to self-renewal and pluripotency (25).…”

Section: E Heparan Sulfate (Hs) On Mouse Es Cells Is Reduced Bymentioning

confidence: 99%

The function of glycan structures expressed on embryonic stem cells

Nishihara

2009

TIGG

Self Cite

View full text Add to dashboard Cite

The identities and amounts of glycan structures present on cell surfaces can change dramatically during development. UndiŠerentiated embryonic stem (ES) cells also express glycan structures on their surfaces and these determine some of the characteristics of the cells. Some of the glycan structures on ES cells are speciˆc to these cells and can be used as markers for ES cells. Others, such as heparan sulfate, are not speciˆc to ES cells but nevertheless have an important function in the cells. In this review, I describe the use of carbohydrate antigens as markers of ES cells and also consider the function of heparan sulfate, a sulfated carbohydrate structure on ES cells, in the maintenance of self-renewal and pluripotency.

show abstract

Proteoglycan UDP-Galactose:β-Xylose β1,4-Galactosyltransferase I Is Essential for Viability inDrosophila melanogaster

Cited by 42 publications

References 36 publications

Molecular Cloning and Identification of 3′-Phosphoadenosine 5′-Phosphosulfate Transporter

Molecular Cloning and Identification of 3′-Phosphoadenosine 5′-Phosphosulfate Transporter

The Twisted Abdomen Phenotype of Drosophila POMT1 and POMT2 Mutants Coincides with Their Heterophilic Protein O-Mannosyltransferase Activity

The function of glycan structures expressed on embryonic stem cells

Contact Info

Product

Resources

About