Localization of three human polypeptide GalNAc-transferases in HeLa cells suggests initiation of O-linked glycosylation throughout the Golgi apparatus.

S Röttger; J White; H H Wandall; J C Olivo; A Stark; E P Bennett; C Whitehouse; E G Berger; H Clausen; T Nilsson

Localization of three human polypeptide GalNAc-transferases in HeLa cells suggests initiation of O-linked glycosylation throughout the Golgi apparatus.

S Röttger, J White, H H Wandall, J C Olivo, A Stark, E P Bennett, C Whitehouse, E G Berger, H Clausen, T Nilsson

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

214 Citationer (Scopus)

Abstract

Udgivelsesdato: 1998-Jan

Originalsprog	Engelsk
Tidsskrift	Journal of Cell Science
Vol/bind	111 ( Pt 1)
Sider (fra-til)	45-60
Antal sider	15
ISSN	0021-9533
Status	Udgivet - 1998

Bibliografisk note

Keywords: Base Sequence; Cell Compartmentation; Cloning, Molecular; Endoplasmic Reticulum; Epitopes; Fluorescent Antibody Technique; Glycosylation; Golgi Apparatus; Hela Cells; Humans; Isoenzymes; Microscopy, Electron; Molecular Sequence Data; N-Acetylgalactosaminyltransferases; Substrate Specificity

Citationsformater

@article{d9fb9b60849d11dd81b0000ea68e967b,

title = "Localization of three human polypeptide GalNAc-transferases in HeLa cells suggests initiation of O-linked glycosylation throughout the Golgi apparatus.",

abstract = "O-glycosylation of proteins is initiated by a family of UDP-N-acetylgalactosamine:polypeptide N-acetylgalactos-aminyltransferases (GalNAc-T). In this study, we have localized endogenous and epitope-tagged human GalNAc-T1, -T2 and -T3 to the Golgi apparatus in HeLa cells by subcellular fractionation, immunofluorescence and immunoelectron microscopy. We show that all three GalNAc-transferases are concentrated about tenfold in Golgi stacks over Golgi associated tubular-vesicular membrane structures. Surprisingly, we find that GalNAc-T1, -T2 and -T3 are present throughout the Golgi stack suggesting that initiation of O-glycosylation may not be restricted to the cis Golgi, but occur at multiple sites within the Golgi apparatus. GalNAc-T1 distributes evenly across the Golgi stack whereas GalNAc-T2 and -T3 reside preferentially on the trans side and in the medial part of the Golgi stack, respectively. Moreover, we have investigated the possibility of O-glycan initiation in pre-Golgi compartments such as the ER. We could not detect endogenous polypeptide GalNAc-transferase activity in the ER of HeLa cells, neither by subcellular fractionation nor by situ glycosylation of an ER-retained form of CD8 (CD8/E19). However, upon relocation of chimeric GalNAc-T1 or -T2 to the ER, CD8/E19 is glycosylated with different efficiencies indicating that all components required for initiation of O-glycosylation are present in the ER except for polypeptide GalNAc-transferases.",

author = "S R{\"o}ttger and J White and Wandall, {H H} and Olivo, {J C} and A Stark and Bennett, {E P} and C Whitehouse and Berger, {E G} and H Clausen and T Nilsson",

note = "Keywords: Base Sequence; Cell Compartmentation; Cloning, Molecular; Endoplasmic Reticulum; Epitopes; Fluorescent Antibody Technique; Glycosylation; Golgi Apparatus; Hela Cells; Humans; Isoenzymes; Microscopy, Electron; Molecular Sequence Data; N-Acetylgalactosaminyltransferases; Substrate Specificity",

year = "1998",

language = "English",

volume = "111 ( Pt 1)",

pages = "45--60",

journal = "Journal of Cell Science",

issn = "0021-9533",

publisher = "The/Company of Biologists Ltd.",

}

TY - JOUR

T1 - Localization of three human polypeptide GalNAc-transferases in HeLa cells suggests initiation of O-linked glycosylation throughout the Golgi apparatus.

AU - Röttger, S

AU - White, J

AU - Wandall, H H

AU - Olivo, J C

AU - Stark, A

AU - Bennett, E P

AU - Whitehouse, C

AU - Berger, E G

AU - Clausen, H

AU - Nilsson, T

N1 - Keywords: Base Sequence; Cell Compartmentation; Cloning, Molecular; Endoplasmic Reticulum; Epitopes; Fluorescent Antibody Technique; Glycosylation; Golgi Apparatus; Hela Cells; Humans; Isoenzymes; Microscopy, Electron; Molecular Sequence Data; N-Acetylgalactosaminyltransferases; Substrate Specificity

PY - 1998

Y1 - 1998

N2 - O-glycosylation of proteins is initiated by a family of UDP-N-acetylgalactosamine:polypeptide N-acetylgalactos-aminyltransferases (GalNAc-T). In this study, we have localized endogenous and epitope-tagged human GalNAc-T1, -T2 and -T3 to the Golgi apparatus in HeLa cells by subcellular fractionation, immunofluorescence and immunoelectron microscopy. We show that all three GalNAc-transferases are concentrated about tenfold in Golgi stacks over Golgi associated tubular-vesicular membrane structures. Surprisingly, we find that GalNAc-T1, -T2 and -T3 are present throughout the Golgi stack suggesting that initiation of O-glycosylation may not be restricted to the cis Golgi, but occur at multiple sites within the Golgi apparatus. GalNAc-T1 distributes evenly across the Golgi stack whereas GalNAc-T2 and -T3 reside preferentially on the trans side and in the medial part of the Golgi stack, respectively. Moreover, we have investigated the possibility of O-glycan initiation in pre-Golgi compartments such as the ER. We could not detect endogenous polypeptide GalNAc-transferase activity in the ER of HeLa cells, neither by subcellular fractionation nor by situ glycosylation of an ER-retained form of CD8 (CD8/E19). However, upon relocation of chimeric GalNAc-T1 or -T2 to the ER, CD8/E19 is glycosylated with different efficiencies indicating that all components required for initiation of O-glycosylation are present in the ER except for polypeptide GalNAc-transferases.

AB - O-glycosylation of proteins is initiated by a family of UDP-N-acetylgalactosamine:polypeptide N-acetylgalactos-aminyltransferases (GalNAc-T). In this study, we have localized endogenous and epitope-tagged human GalNAc-T1, -T2 and -T3 to the Golgi apparatus in HeLa cells by subcellular fractionation, immunofluorescence and immunoelectron microscopy. We show that all three GalNAc-transferases are concentrated about tenfold in Golgi stacks over Golgi associated tubular-vesicular membrane structures. Surprisingly, we find that GalNAc-T1, -T2 and -T3 are present throughout the Golgi stack suggesting that initiation of O-glycosylation may not be restricted to the cis Golgi, but occur at multiple sites within the Golgi apparatus. GalNAc-T1 distributes evenly across the Golgi stack whereas GalNAc-T2 and -T3 reside preferentially on the trans side and in the medial part of the Golgi stack, respectively. Moreover, we have investigated the possibility of O-glycan initiation in pre-Golgi compartments such as the ER. We could not detect endogenous polypeptide GalNAc-transferase activity in the ER of HeLa cells, neither by subcellular fractionation nor by situ glycosylation of an ER-retained form of CD8 (CD8/E19). However, upon relocation of chimeric GalNAc-T1 or -T2 to the ER, CD8/E19 is glycosylated with different efficiencies indicating that all components required for initiation of O-glycosylation are present in the ER except for polypeptide GalNAc-transferases.

M3 - Journal article

C2 - 9394011

SN - 0021-9533

VL - 111 ( Pt 1)

SP - 45

EP - 60

JO - Journal of Cell Science

JF - Journal of Cell Science

ER -