Influence of N-glycosylation on Saccharomyces cerevisiae morphology: a golgi glycosylation mutant shows cell division defects.
The N-glycosylation mutants (mnn1 and mnn1 och1) show different morphological characteristics at the restrictive and nonpermissive temperature. We deleted the MNN1 to eliminate the terminal alpha1, 3-linked mannose of hypermannosylation and deleted the OCH1 to block the elongation of the main backbone chain. The mnn1 cells exhibited no observable change with respect to the wild-type strain at 28 degrees C and 37 degrees C, but the mnn1 och1 double mutant exhibited defects in cell cytokinesis, showed a slower growth rate, and became temperature-sensitive. Meanwhile, the mnn1 och1 mutant tended to aggregate, which was probably due to the glycolsylation defect. Loss of mannosyl-phosphate-accepting sites in this mutant migth result in reduced charge repulsion between cell surfaces. Pyridylaminated glycans were profiled and purified through an NH(2) column by size-fractionation high-performance liquid chromatography. Matrix assisted laser desoption/ionization time of flight mass spectrometry (MALDI TOF/MS) analysis of the N-glycan structure of the mnn1 och1 mutant revealed that the main component is Man(8)GlcNAc(2).
Zhou
Jungang
J
State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, 250100, Jinan, People's Republic of China.
Zhang
Houcheng
H
Liu
Xianwei
X
Wang
Peng George
PG
Qi
Qingsheng
Q
D002455:Cell Division / Q000235:genetics*; D006031:Glycosylation; D006056:Golgi Apparatus / Q000378:metabolism; D008364:Mannosyltransferases / Q000235:genetics; D008562:Membrane Glycoproteins / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
D008562:Membrane Glycoproteins; C075636:OCH1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; C086604:MNN1 protein, S cerevisiae; D008364:Mannosyltransferases