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).
State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, 250100, Jinan, People's Republic of China.
Jungang
J
Zhou
Houcheng
H
Zhang
Xianwei
X
Liu
Peng George
PG
Wang
Qingsheng
Q
Qi
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