Reduction in Golgi apparatus dimension in the absence of a residential protein, N-acetylglucosaminyltransferase v

Zhizhong Dong, Christian Zuber, Michael Pierce, Pamela Stanley, Jürgen Roth

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Various proteins are involved in the generation and maintenance of the membrane complex known as the Golgi apparatus. We have used mutant Chinese hamster ovary (CHO) cell lines Lec4 and Lec4A lacking N- acetylglucosaminyltransferase V (GlcNAcT-V, MGAT5) activity and protein in the Golgi apparatus to study the effects of the absence of a single glycosyltransferase on the Golgi apparatus dimension. Quantification of immunofluorescence in serial confocal sections for Golgi α-mannosidase II and electron microscopic morphometry revealed a reduction in Golgi volume density up to 49 % in CHO Lec4 and CHO Lec4A cells compared to parental CHO cells. This reduction in Golgi volume density could be reversed by stable transfection of Lec4 cells with a cDNA encoding Mgat5. Inhibition of the synthesis of β1,6-branched N-glycans by swainsonine had no effect on Golgi volume density. In addition, no effect on Golgi volume density was observed in CHO Lec1 cells that contain enzymatically active GlcNAcT-V, but cannot synthesize β1,6-branched glycans due to an inactive GlcNAcT-I in their Golgi apparatus. These results indicate that it may be the absence of the GlcNAcT-V protein that is the determining factor in reducing Golgi volume density. No dimensional differences existed in cross-sectioned cisternal stacks between Lec4 and control CHO cells, but significantly reduced Golgi stack hits were observed in cross-sectioned Lec4 cells. Therefore, the Golgi apparatus dimensional change in Lec4 and Lec4A cells may be due to a compaction of the organelle.

Original languageEnglish (US)
Pages (from-to)153-164
Number of pages12
JournalHistochemistry and Cell Biology
Volume141
Issue number2
DOIs
StatePublished - Feb 2014

Fingerprint

Golgi Apparatus
Cricetulus
Ovary
Proteins
alpha-1,6-mannosylglycoprotein beta 1,6-N-acetylglucosaminyltransferase
Polysaccharides
Swainsonine
Glycosyltransferases
N-acetyllactosaminide beta-1,6-N-acetylglucosaminyltransferase
Organelles
Fluorescent Antibody Technique
Transfection
Complementary DNA
Maintenance
Electrons
Cell Line
Membranes

Keywords

  • β1,6-branched N-glycans
  • CHO Lec1 cells
  • CHO Lec4 cells
  • Golgi α-mannosidase II
  • Golgi apparatus
  • Golgi volume density
  • N- acetylglucosaminyltransferase I
  • N-acetylglucosaminyltransferase V

ASJC Scopus subject areas

  • Cell Biology
  • Histology
  • Medical Laboratory Technology
  • Molecular Biology

Cite this

Reduction in Golgi apparatus dimension in the absence of a residential protein, N-acetylglucosaminyltransferase v. / Dong, Zhizhong; Zuber, Christian; Pierce, Michael; Stanley, Pamela; Roth, Jürgen.

In: Histochemistry and Cell Biology, Vol. 141, No. 2, 02.2014, p. 153-164.

Research output: Contribution to journalArticle

Dong, Zhizhong ; Zuber, Christian ; Pierce, Michael ; Stanley, Pamela ; Roth, Jürgen. / Reduction in Golgi apparatus dimension in the absence of a residential protein, N-acetylglucosaminyltransferase v. In: Histochemistry and Cell Biology. 2014 ; Vol. 141, No. 2. pp. 153-164.
@article{d2aa03be387642ce8c7265a5f140a7b2,
title = "Reduction in Golgi apparatus dimension in the absence of a residential protein, N-acetylglucosaminyltransferase v",
abstract = "Various proteins are involved in the generation and maintenance of the membrane complex known as the Golgi apparatus. We have used mutant Chinese hamster ovary (CHO) cell lines Lec4 and Lec4A lacking N- acetylglucosaminyltransferase V (GlcNAcT-V, MGAT5) activity and protein in the Golgi apparatus to study the effects of the absence of a single glycosyltransferase on the Golgi apparatus dimension. Quantification of immunofluorescence in serial confocal sections for Golgi α-mannosidase II and electron microscopic morphometry revealed a reduction in Golgi volume density up to 49 {\%} in CHO Lec4 and CHO Lec4A cells compared to parental CHO cells. This reduction in Golgi volume density could be reversed by stable transfection of Lec4 cells with a cDNA encoding Mgat5. Inhibition of the synthesis of β1,6-branched N-glycans by swainsonine had no effect on Golgi volume density. In addition, no effect on Golgi volume density was observed in CHO Lec1 cells that contain enzymatically active GlcNAcT-V, but cannot synthesize β1,6-branched glycans due to an inactive GlcNAcT-I in their Golgi apparatus. These results indicate that it may be the absence of the GlcNAcT-V protein that is the determining factor in reducing Golgi volume density. No dimensional differences existed in cross-sectioned cisternal stacks between Lec4 and control CHO cells, but significantly reduced Golgi stack hits were observed in cross-sectioned Lec4 cells. Therefore, the Golgi apparatus dimensional change in Lec4 and Lec4A cells may be due to a compaction of the organelle.",
keywords = "β1,6-branched N-glycans, CHO Lec1 cells, CHO Lec4 cells, Golgi α-mannosidase II, Golgi apparatus, Golgi volume density, N- acetylglucosaminyltransferase I, N-acetylglucosaminyltransferase V",
author = "Zhizhong Dong and Christian Zuber and Michael Pierce and Pamela Stanley and J{\"u}rgen Roth",
year = "2014",
month = "2",
doi = "10.1007/s00418-013-1146-1",
language = "English (US)",
volume = "141",
pages = "153--164",
journal = "Histochemistry and Cell Biology",
issn = "0948-6143",
publisher = "Springer Verlag",
number = "2",

}

TY - JOUR

T1 - Reduction in Golgi apparatus dimension in the absence of a residential protein, N-acetylglucosaminyltransferase v

AU - Dong, Zhizhong

AU - Zuber, Christian

AU - Pierce, Michael

AU - Stanley, Pamela

AU - Roth, Jürgen

PY - 2014/2

Y1 - 2014/2

N2 - Various proteins are involved in the generation and maintenance of the membrane complex known as the Golgi apparatus. We have used mutant Chinese hamster ovary (CHO) cell lines Lec4 and Lec4A lacking N- acetylglucosaminyltransferase V (GlcNAcT-V, MGAT5) activity and protein in the Golgi apparatus to study the effects of the absence of a single glycosyltransferase on the Golgi apparatus dimension. Quantification of immunofluorescence in serial confocal sections for Golgi α-mannosidase II and electron microscopic morphometry revealed a reduction in Golgi volume density up to 49 % in CHO Lec4 and CHO Lec4A cells compared to parental CHO cells. This reduction in Golgi volume density could be reversed by stable transfection of Lec4 cells with a cDNA encoding Mgat5. Inhibition of the synthesis of β1,6-branched N-glycans by swainsonine had no effect on Golgi volume density. In addition, no effect on Golgi volume density was observed in CHO Lec1 cells that contain enzymatically active GlcNAcT-V, but cannot synthesize β1,6-branched glycans due to an inactive GlcNAcT-I in their Golgi apparatus. These results indicate that it may be the absence of the GlcNAcT-V protein that is the determining factor in reducing Golgi volume density. No dimensional differences existed in cross-sectioned cisternal stacks between Lec4 and control CHO cells, but significantly reduced Golgi stack hits were observed in cross-sectioned Lec4 cells. Therefore, the Golgi apparatus dimensional change in Lec4 and Lec4A cells may be due to a compaction of the organelle.

AB - Various proteins are involved in the generation and maintenance of the membrane complex known as the Golgi apparatus. We have used mutant Chinese hamster ovary (CHO) cell lines Lec4 and Lec4A lacking N- acetylglucosaminyltransferase V (GlcNAcT-V, MGAT5) activity and protein in the Golgi apparatus to study the effects of the absence of a single glycosyltransferase on the Golgi apparatus dimension. Quantification of immunofluorescence in serial confocal sections for Golgi α-mannosidase II and electron microscopic morphometry revealed a reduction in Golgi volume density up to 49 % in CHO Lec4 and CHO Lec4A cells compared to parental CHO cells. This reduction in Golgi volume density could be reversed by stable transfection of Lec4 cells with a cDNA encoding Mgat5. Inhibition of the synthesis of β1,6-branched N-glycans by swainsonine had no effect on Golgi volume density. In addition, no effect on Golgi volume density was observed in CHO Lec1 cells that contain enzymatically active GlcNAcT-V, but cannot synthesize β1,6-branched glycans due to an inactive GlcNAcT-I in their Golgi apparatus. These results indicate that it may be the absence of the GlcNAcT-V protein that is the determining factor in reducing Golgi volume density. No dimensional differences existed in cross-sectioned cisternal stacks between Lec4 and control CHO cells, but significantly reduced Golgi stack hits were observed in cross-sectioned Lec4 cells. Therefore, the Golgi apparatus dimensional change in Lec4 and Lec4A cells may be due to a compaction of the organelle.

KW - β1,6-branched N-glycans

KW - CHO Lec1 cells

KW - CHO Lec4 cells

KW - Golgi α-mannosidase II

KW - Golgi apparatus

KW - Golgi volume density

KW - N- acetylglucosaminyltransferase I

KW - N-acetylglucosaminyltransferase V

UR - http://www.scopus.com/inward/record.url?scp=84893863845&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84893863845&partnerID=8YFLogxK

U2 - 10.1007/s00418-013-1146-1

DO - 10.1007/s00418-013-1146-1

M3 - Article

VL - 141

SP - 153

EP - 164

JO - Histochemistry and Cell Biology

JF - Histochemistry and Cell Biology

SN - 0948-6143

IS - 2

ER -