naa a22 4500 569238005 CHVBK 20190724065209.0 cr unu---uuuuu 190724e20140201xx s 000 0 eng 10.1007/s00418-013-1146-1 doi 10.1007/s00418-013-1146-1 doi (NATIONALLICENCE)springer-10.1007/s00418-013-1146-1 (ZORA)oai:www.zora.uzh.ch:172012 570 ddc 610 ddc Reduction in Golgi apparatus dimension in the absence of a residential protein, N -acetylglucosaminyltransferase V [Elektronische Daten] [Zhizhong Dong, Christian Zuber, Michael Pierce, Pamela Stanley, Jürgen Roth] openAccess eu-repo 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. Springer-Verlag Berlin Heidelberg, 2013 Golgi apparatus nationallicence N -acetylglucosaminyltransferase V nationallicence N -acetylglucosaminyltransferase I nationallicence CHO Lec4 cells nationallicence CHO Lec1 cells nationallicence β1,6-branched N -glycans nationallicence Golgi volume density nationallicence Golgi α-mannosidase II nationallicence GlcNAcT-I : N-acetylglucosaminyltransferase I nationallicence GlcNAcT-V, MGAT5 : N-acetylglucosaminyltransferase V nationallicence CHO : Chinese hamster ovary cells nationallicence GlcNAc : N-acetylglucosamine nationallicence Man : Mannose nationallicence L-PHA : Leukoagglutinating Phaseolus vulgaris lectin nationallicence Dig : Digoxigenin nationallicence 142-005 zora Cell Biology, Molecular Biology, Medical Laboratory Technology, Histology zora Dong Zhizhong Division of Cell and Molecular Pathology, Department of Pathology, University of Zürich, 8091, Zürich, Switzerland aut Zuber Christian Division of Cell and Molecular Pathology, Department of Pathology, University of Zürich, 8091, Zürich, Switzerland aut Pierce Michael Complex Carbohydrate Research Center, Department of Biochemistry and Molecular Biology, University of Georgia, 30602, Athens, GA, USA aut Stanley Pamela Department of Cell Biology, Albert Einstein College of Medicine, 10461, New York, NY, USA aut Roth Jürgen Division of Cell and Molecular Pathology, Department of Pathology, University of Zürich, 8091, Zürich, Switzerland aut Histochemistry and Cell Biology Springer Berlin Heidelberg 141/2(2014-02-01), 153-164 0948-6143 141:2<153 2014 141 418 https://doi.org/10.1007/s00418-013-1146-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00418-013-1146-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Dong Zhizhong Division of Cell and Molecular Pathology, Department of Pathology, University of Zürich, 8091, Zürich, Switzerland aut NATIONALLICENCE 700 1- Zuber Christian Division of Cell and Molecular Pathology, Department of Pathology, University of Zürich, 8091, Zürich, Switzerland aut NATIONALLICENCE 700 1- Pierce Michael Complex Carbohydrate Research Center, Department of Biochemistry and Molecular Biology, University of Georgia, 30602, Athens, GA, USA aut NATIONALLICENCE 700 1- Stanley Pamela Department of Cell Biology, Albert Einstein College of Medicine, 10461, New York, NY, USA aut NATIONALLICENCE 700 1- Roth Jürgen Division of Cell and Molecular Pathology, Department of Pathology, University of Zürich, 8091, Zürich, Switzerland aut NATIONALLICENCE 773 0- Histochemistry and Cell Biology Springer Berlin Heidelberg 141/2(2014-02-01), 153-164 0948-6143 141:2<153 2014 141 418 ZORA 856 40 https://www.zora.uzh.ch/id/eprint/172012/1/ZORA_NL_172012.pdf text/html WWW-Backlink auf das Repository ZORA 700 1- Dong Zhizhong joint author ZORA 700 1- Zuber Christian joint author ZORA 700 1- Pierce Michael joint author ZORA 700 1- Stanley Pamela joint author ZORA 700 1- Roth Jürgen joint author ZORA 773 0- Histochemistry and Cell Biology 141(2):153-164 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer ZORA ZORA ZORA Journal Article openAccess