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Knockout of RSN1, TVP18 or CSC1‐2 causes perturbation of Golgi cisternae in Pichia pastoris

Author
Organization
Abstract
The structural organization of the Golgi stacks in mammalian cells is intrinsically linked to function, including glycosylation, but the role of morphology is less clear in lower eukaryotes. Here we investigated the link between the structural organization of the Golgi and secretory pathway function using Pichia pastoris as a model system. To unstack the Golgi cisternae, we disrupted 18 genes encoding proteins in the secretory pathway without loss of viability. Using biosensors, confocal microscopy and transmission electron microscopy we identified three strains with irreversible perturbations in the stacking of the Golgi cisternae, all of which had disruption in genes that encode proteins with annotated function as or homology to calcium/calcium permeable ion channels. Despite this, no variation in the secretory pathway for ER size, whole cell glycomics or recombinant protein glycans was observed. Our investigations showed the robust nature of the secretory pathway in P. pastoris and suggest that Ca2+ concentration, homeostasis or signalling may play a significant role for Golgi stacking in this organism and should be investigated in other organisms.
Keywords
Genetics, Cell Biology, Biochemistry, Molecular Biology, Structural Biology, glycosylation, Golgi, Pichia pastoris, Komagataella phaffi, stacked cisternae, secretory pathway, CIS-GOLGI, PROTEINS, IDENTIFICATION, GLYCOSYLATION, ORGANIZATION, TRAFFICKING, SELECTION, STACKING, HOMOLOG, COMPLEX

Citation

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MLA
Aw, Rochelle, et al. “Knockout of RSN1, TVP18 or CSC1‐2 Causes Perturbation of Golgi Cisternae in Pichia Pastoris.” TRAFFIC, 2020, doi:10.1111/tra.12773.
APA
Aw, R., De Wachter, C., Laukens, B., De Rycke, R., De Bruyne, M., Bell, D., … Polizzi, K. M. (2020). Knockout of RSN1, TVP18 or CSC1‐2 causes perturbation of Golgi cisternae in Pichia pastoris. TRAFFIC. https://doi.org/10.1111/tra.12773
Chicago author-date
Aw, Rochelle, Charlot De Wachter, Bram Laukens, Riet De Rycke, Michiel De Bruyne, David Bell, Nico Callewaert, and Karen M. Polizzi. 2020. “Knockout of RSN1, TVP18 or CSC1‐2 Causes Perturbation of Golgi Cisternae in Pichia Pastoris.” TRAFFIC. https://doi.org/10.1111/tra.12773.
Chicago author-date (all authors)
Aw, Rochelle, Charlot De Wachter, Bram Laukens, Riet De Rycke, Michiel De Bruyne, David Bell, Nico Callewaert, and Karen M. Polizzi. 2020. “Knockout of RSN1, TVP18 or CSC1‐2 Causes Perturbation of Golgi Cisternae in Pichia Pastoris.” TRAFFIC. doi:10.1111/tra.12773.
Vancouver
1.
Aw R, De Wachter C, Laukens B, De Rycke R, De Bruyne M, Bell D, et al. Knockout of RSN1, TVP18 or CSC1‐2 causes perturbation of Golgi cisternae in Pichia pastoris. TRAFFIC. 2020;
IEEE
[1]
R. Aw et al., “Knockout of RSN1, TVP18 or CSC1‐2 causes perturbation of Golgi cisternae in Pichia pastoris,” TRAFFIC, 2020.
@article{8693843,
  abstract     = {The structural organization of the Golgi stacks in mammalian cells is intrinsically linked to function, including glycosylation, but the role of morphology is less clear in lower eukaryotes. Here we investigated the link between the structural organization of the Golgi and secretory pathway function using Pichia pastoris as a model system. To unstack the Golgi cisternae, we disrupted 18 genes encoding proteins in the secretory pathway without loss of viability. Using biosensors, confocal microscopy and transmission electron microscopy we identified three strains with irreversible perturbations in the stacking of the Golgi cisternae, all of which had disruption in genes that encode proteins with annotated function as or homology to calcium/calcium permeable ion channels. Despite this, no variation in the secretory pathway for ER size, whole cell glycomics or recombinant protein glycans was observed. Our investigations showed the robust nature of the secretory pathway in P. pastoris and suggest that Ca2+ concentration, homeostasis or signalling may play a significant role for Golgi stacking in this organism and should be investigated in other organisms.},
  author       = {Aw, Rochelle and De Wachter, Charlot and Laukens, Bram and De Rycke, Riet and De Bruyne, Michiel and Bell, David and Callewaert, Nico and Polizzi, Karen M.},
  issn         = {1398-9219},
  journal      = {TRAFFIC},
  keywords     = {Genetics,Cell Biology,Biochemistry,Molecular Biology,Structural Biology,glycosylation,Golgi,Pichia pastoris,Komagataella phaffi,stacked cisternae,secretory pathway,CIS-GOLGI,PROTEINS,IDENTIFICATION,GLYCOSYLATION,ORGANIZATION,TRAFFICKING,SELECTION,STACKING,HOMOLOG,COMPLEX},
  language     = {eng},
  pages        = {16},
  title        = {Knockout of RSN1, TVP18 or CSC1‐2 causes perturbation of Golgi cisternae in Pichia pastoris},
  url          = {http://dx.doi.org/10.1111/tra.12773},
  year         = {2020},
}

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