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PMID:19139268

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Citation

Johswich, A, Kraft, B, Wuhrer, M, Berger, M, Deelder, AM, Hokke, CH, Gerardy-Schahn, R and Bakker, H (2009) Golgi targeting of Drosophila melanogaster beta4GalNAcTB requires a DHHC protein family-related protein as a pilot. J. Cell Biol. 184:173-83

Abstract

Drosophila melanogaster beta4GalNAcTB mutant flies revealed that this particular N-acetylgalactosaminyltransferase is predominant in the formation of lacdiNAc (GalNAcbeta1,4GlcNAc)-modified glycolipids, but enzymatic activity could not be confirmed for the cloned enzyme. Using a heterologous expression cloning approach, we isolated beta4GalNAcTB together with beta4GalNAcTB pilot (GABPI), a multimembrane-spanning protein related to Asp-His-His-Cys (DHHC) proteins but lacking the DHHC consensus sequence. In the absence of GABPI, inactive beta4GalNAcTB is trapped in the endoplasmic reticulum (ER). Coexpression of beta4GalNAcTB and GABPI generates the active enzyme that is localized together with GABPI in the Golgi. GABPI associates with beta4GalNAcTB and, when expressed with an ER retention signal, holds active beta4GalNAcTB in the ER. Importantly, treatment of isolated membrane vesicles with Triton X-100 disturbs beta4GalNAcTB activity. This phenomenon occurs with multimembrane-spanning glycosyltransferases but is normally not a property of glycosyltransferases with one membrane anchor. In summary, our data provide evidence that GABPI is required for ER export and activity of beta4GalNAcTB.

Links

PubMed PMC2615082 Online version:10.1083/jcb.200801071

Keywords

Animals; CHO Cells; Cell Line; Cloning, Molecular; Cricetinae; Cricetulus; Drosophila Proteins/chemistry; Drosophila Proteins/metabolism; Drosophila melanogaster/metabolism; Endoplasmic Reticulum/metabolism; Gene Library; Glycolipids/metabolism; Glycosylation; Golgi Apparatus/metabolism; Humans; Lactose/analogs & derivatives; Lactose/metabolism; Membrane Proteins/metabolism; N-Acetylgalactosaminyltransferases/chemistry; N-Acetylgalactosaminyltransferases/metabolism; Octoxynol/pharmacology; Protein Interaction Mapping; Protein Transport; Saponins/pharmacology

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

DROME:Q7KN92

involved_in

GO:0009247: glycolipid biosynthetic process

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:Q7KN92

enables

GO:0033842: N-acetyl-beta-glucosaminyl-glycoprotein 4-beta-N-acetylgalactosaminyltransferase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

DROME:Q7KN92

located_in

GO:0005794: Golgi apparatus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:Q86NU9

involved_in

GO:0009247: glycolipid biosynthetic process

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:Q86NU9

enables

GO:0033842: N-acetyl-beta-glucosaminyl-glycoprotein 4-beta-N-acetylgalactosaminyltransferase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

DROME:Q86NU9

located_in

GO:0005794: Golgi apparatus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:Q9VAQ8

involved_in

GO:0009247: glycolipid biosynthetic process

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:Q9VAQ8

enables

GO:0033842: N-acetyl-beta-glucosaminyl-glycoprotein 4-beta-N-acetylgalactosaminyltransferase activity

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete

DROME:Q9VAQ8

located_in

GO:0005794: Golgi apparatus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:Q9VQK9

involved_in

GO:0006888: endoplasmic reticulum to Golgi vesicle-mediated transport

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:Q9VQK9

located_in

GO:0005794: Golgi apparatus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete

DROME:X2J6Q6

involved_in

GO:0006888: endoplasmic reticulum to Golgi vesicle-mediated transport

ECO:0000314: direct assay evidence used in manual assertion

P

Seeded From UniProt

complete

DROME:X2J6Q6

located_in

GO:0005794: Golgi apparatus

ECO:0000314: direct assay evidence used in manual assertion

C

Seeded From UniProt

complete


See also

References

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