scholarly journals Semi-Intact Cell Systems - Application to the Analysis of Membrane Trafficking Between the Endoplasmic Reticulum and the Golgi Apparatus and of Cell Cycle-Dependent Changes in the Morphology of These Organelles

10.5772/31440 ◽  
2012 ◽  
Author(s):  
Masayuki Murata ◽  
Fumi Kano
Keyword(s):  
Cell Cycle ◽  
Endoplasmic Reticulum ◽  
Golgi Apparatus ◽  
Membrane Trafficking ◽  
Intact Cell ◽  
Cell Systems ◽  
Cycle Dependent

scholarly journals Cell cycle–dependent phosphorylation of Sec4p controls membrane deposition during cytokinesis

2016 ◽  
Vol 214 (6) ◽  
pp. 691-703 ◽  
Author(s):  
Dante Lepore ◽  
Olya Spassibojko ◽  
Gabrielle Pinto ◽  
Ruth N. Collins
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Cycle ◽  
Membrane Trafficking ◽  
Intracellular Trafficking ◽  
Rab Gtpase ◽  
Rab Gtpases ◽  
Positive Regulator ◽  
Physiological Relevance ◽  
A Cell ◽  
Cycle Dependent

Intracellular trafficking is an essential and conserved eukaryotic process. Rab GTPases are a family of proteins that regulate and provide specificity for discrete membrane trafficking steps by harnessing a nucleotide-bound cycle. Global proteomic screens have revealed many Rab GTPases as phosphoproteins, but the effects of this modification are not well understood. Using the Saccharomyces cerevisiae Rab GTPase Sec4p as a model, we have found that phosphorylation negatively regulates Sec4p function by disrupting the interaction with the exocyst complex via Sec15p. We demonstrate that phosphorylation of Sec4p is a cell cycle–dependent process associated with cytokinesis. Through a genomic kinase screen, we have also identified the polo-like kinase Cdc5p as a positive regulator of Sec4p phosphorylation. Sec4p spatially and temporally localizes with Cdc5p exclusively when Sec4p phosphorylation levels peak during the cell cycle, indicating Sec4p is a direct Cdc5p substrate. Our data suggest the physiological relevance of Sec4p phosphorylation is to facilitate the coordination of membrane-trafficking events during cytokinesis.

2P192 Cell cycle-dependent dynamics of the endoplasmic reticulum (ER) in mammalian cells (I): Mechanism of disruption of the ER network

2004 ◽  
Vol 44 (supplement) ◽  
pp. S157
Author(s):  
A.R. Tanaka ◽  
P. Kano ◽  
H. Kondo ◽  
S. Yamauchi ◽  
N. Hosokawa ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Endoplasmic Reticulum ◽  
Mammalian Cells ◽  
Cycle Dependent

scholarly journals The proteasomal deubiquitinating enzyme PSMD14 regulates macroautophagy by controlling Golgi-to-ER retrograde transport

2020 ◽  
Author(s):  
HA Bustamante ◽  
K Cereceda ◽  
AE González ◽  
GE Valenzuela ◽  
Y Cheuquemilla ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Golgi Apparatus ◽  
Membrane Trafficking ◽  
Retrograde Transport ◽  
Deubiquitinating Enzyme ◽  
20S Proteasome ◽  
Biological Processes ◽  
Pharmacological Inhibition ◽  
Protein Membrane ◽  
A Subunit

ABSTRACTUbiquitination regulates several biological processes. Here, we search for ubiquitin-related genes implicated in protein membrane trafficking performing a High-Content siRNA Screening including 1,187 genes of the human “ubiquitinome” using Amyloid Precursor Protein (APP) as a reporter. We identified the deubiquitinating enzyme PSMD14, a subunit of the 19S regulatory particle of the proteasome, specific for K63-Ub chains in cells, as a novel key regulator of Golgi-to-endoplasmic reticulum (ER) retrograde transport. Silencing or pharmacological inhibition of PSMD14 caused a robust and rapid inhibition of Golgi-to-ER retrograde transport which leads to a potent blockage of macroautophagy by a mechanism associated with the retention of Atg9A and Rab1A at the Golgi apparatus. Because pharmacological inhibition of the proteolytic core of the 20S proteasome did not recapitulate these effects, we concluded that PSMD14, and their K-63-Ub chains, act as a crucial regulator factor for macroautophagy by controlling Golgi-to-ER retrograde transport.

scholarly journals Reconstitution of cell cycle-dependent morphological chanbes of the endoplasmic reticulum in semi-intact CHO cells

2001 ◽  
Vol 41 (supplement) ◽  
pp. S211
Author(s):  
M. Murata ◽  
F. Kano ◽  
H. Kondo ◽  
N. Hosokawa ◽  
K. Nagata
Keyword(s):  
Cell Cycle ◽  
Endoplasmic Reticulum ◽  
Cho Cells ◽  
Cycle Dependent

scholarly journals 2P193 Cell cycle-dependent dynamics of the endoplasmic reticulum (ER) in mammalian cells (II) : Mechanism of membrane fusion of the ER

2004 ◽  
Vol 44 (supplement) ◽  
pp. S158
Author(s):  
F. Kano ◽  
H. Kondo ◽  
S. Yamauchi ◽  
A.R. Tanaka ◽  
N. Hosokawa ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Endoplasmic Reticulum ◽  
Membrane Fusion ◽  
Mammalian Cells ◽  
Cycle Dependent

scholarly journals Golgi duplication in Trypanosoma brucei

2004 ◽  
Vol 165 (3) ◽  
pp. 313-321 ◽  
Author(s):  
Cynthia Y. He ◽  
Helen H. Ho ◽  
Joerg Malsam ◽  
Cecile Chalouni ◽  
Christopher M. West ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Endoplasmic Reticulum ◽  
Golgi Apparatus ◽  
Trypanosoma Brucei ◽  
Basal Body ◽  
Time Course ◽  
Matrix Protein ◽  
De Novo ◽  
Protozoan Parasite ◽  
Er Export

Duplication of the single Golgi apparatus in the protozoan parasite Trypanosoma brucei has been followed by tagging a putative Golgi enzyme and a matrix protein with variants of GFP. Video microscopy shows that the new Golgi appears de novo, near to the old Golgi, about two hours into the cell cycle and grows over a two-hour period until it is the same size as the old Golgi. Duplication of the endoplasmic reticulum (ER) export site follows exactly the same time course. Photobleaching experiments show that the new Golgi is not the exclusive product of the new ER export site. Rather, it is supplied, at least in part, by material directly from the old Golgi. Pharmacological experiments show that the site of the new Golgi and ER export is determined by the location of the new basal body.

scholarly journals Cell Cycle-dependent Regulation of Structure of Endoplasmic Reticulum and Inositol 1,4,5-Trisphosphate-induced Ca2+ Release in Mouse Oocytes and Embryos

2003 ◽  
Vol 14 (1) ◽  
pp. 288-301 ◽  
Author(s):  
Greg FitzHarris ◽  
Petros Marangos ◽  
John Carroll
Keyword(s):  
Cell Cycle ◽  
Endoplasmic Reticulum ◽  
Mitotic Spindle ◽  
Mitotic Division ◽  
Cyclin B ◽  
Dependent Manner ◽  
Inositol 1,4,5 Trisphosphate ◽  
A Cell ◽  
Cycle Dependent ◽  
Mouse Eggs

The organization of endoplasmic reticulum (ER) was examined in mouse eggs undergoing fertilization and in embryos during the first cell cycle. The ER in meiosis II (MII)-arrested mouse eggs is characterized by accumulations (clusters) that are restricted to the cortex of the vegetal hemisphere of the egg. Monitoring ER structure with DiI18 after egg activation has demonstrated that ER clusters disappear at the completion of meiosis II. The ER clusters can be maintained by inhibiting the decrease in cdk1-cyclin B activity by using the proteasome inhibitor MG132, or by microinjecting excess cyclin B. A role for cdk1-cyclin B in ER organization is further suggested by the finding that the cdk inhibitor roscovitine causes the loss of ER clusters in MII eggs. Cortical clusters are specific to meiosis as they do not return in the first mitotic division; rather, the ER aggregates around the mitotic spindle. Inositol 1,4,5-trisphosphate-induced Ca2+ release is also regulated in a cell cycle-dependent manner where it is increased in MII and in the first mitosis. The cell cycle dependent effects on ER structure and inositol 1,4,5-trisphosphate-induced Ca2+ release have implications for understanding meiotic and mitotic control of ER structure and inheritance, and of the mechanisms regulating mitotic Ca2+signaling.

scholarly journals Ultrastructure of cells after reversible dark-induced blocking of mitotic divisions in antheridial filaments of Chara vulgaris L.

2014 ◽  
Vol 49 (3) ◽  
pp. 169-186 ◽  
Author(s):  
Maria Kwiatkowska ◽  
Janusz Maszewski ◽  
Maria M. Maszewska
Keyword(s):  
Cell Cycle ◽  
Endoplasmic Reticulum ◽  
Spatial Distribution ◽  
Cell Growth ◽  
Golgi Apparatus ◽  
Lipid Droplets ◽  
Continuous Darkness ◽  
Mean Size ◽  
Dense Matrices ◽  
The Mean

As compared with the control plants cultured under photoperiodic L : D = =14 : 10 conditions (K w i a t k o w s k a, M a s z e w s k i, 1978), the ultrastructure of nuclei -in cells blocked by a 5 day exposure to continuous darkness is characterized by homogenous arrangement. This homogeneity is maintained in all generations of antheridial filaments irrespective of cell length, which in the controls, being directly correlated with particular type of nuclear structure, may serve as a precise indicator of a given stage of interphase. From similarities in both the spatial distribution and content of condensed chromatin in is concluded that the block of the cell cycle is imposed at the beginning of the G<sub>2</sub> phase. On comparing these cells with the early G<sub>2</sub> period (stage VII) in the control plants, marked changes in the structure of nucleoli were found. They decrease in size by half owing to the complete decline of granular component. The area occupied by endoplasmic reticulum undergoes a 50% reduction. The decrease in the activity of Golgi apparatus expressed by a drop in number of smooth vesicles surrounding a single dictyosome is found to parallel the limited rate of cell growth. The number of coated vesicles and cisterns of dictyosome slightly increases. Mitochondria show typical condensed configuration with dense matrices and swollen cristae, while in the control orthodox forms are prevailing. The mean size of mitochondria is smaller, but their number exceeds that of the control plants. The surface area of mitochondrial profiles is found to remain constant proportion of the cytoplasm section, e.g., about 3%. Dark-cultured antheridial filaments show absolute decline of lipid droplets. No differences were found in structure of plastids and vacuols, as well as in number of ribosomes in cytoplasm surface unit.

scholarly journals The mammalian homolog of yeast Sec13p is enriched in the intermediate compartment and is essential for protein transport from the endoplasmic reticulum to the Golgi apparatus.

1997 ◽  
Vol 17 (1) ◽  
pp. 256-266 ◽  
Author(s):  
B L Tang ◽  
F Peter ◽  
J Krijnse-Locker ◽  
S H Low ◽  
G Griffiths ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Endoplasmic Reticulum ◽  
Golgi Apparatus ◽  
Intact Cell ◽  
Predicted Amino Acid Sequence ◽  
Restrictive Temperature ◽  
Dependent Manner ◽  
Yeast Saccharomyces Cerevisiae ◽  
Golgi Transport ◽  
Intermediate Compartment

The role of COPII components in endoplasmic reticulum (ER)-Golgi transport, first identified in the yeast Saccharomyces cerevisiae, has yet to be fully characterized in higher eukaryotes. A human cDNA whose predicted amino acid sequence showed 70% similarity to the yeast Sec13p has previously been cloned. Antibodies raised against the human SEC13 protein (mSEC13) recognized a cellular protein of 35 kDa in both the soluble and membrane fractions. Like the yeast Sec13p, mSEC13 exist in the cytosol in both monomeric and higher-molecular-weight forms. Immunofluorescence microscopy localized mSEC13 to the characteristic spotty ER-Golgi intermediate compartment (ERGIC) in cells of all species examined, where it colocalized well with the KDEL receptor, an ERGIC marker, at 15 degrees C. Immunoelectron microscopy also localized mSEC13 to membrane structures close to the Golgi apparatus. mSEC13 is essential for ER-to-Golgi transport, since both the His6-tagged mSEC13 recombinant protein and the affinity-purified mSEC13 antibody inhibited the transport of restrictive temperature-arrested vesicular stomatitis virus G protein from the ER to the Golgi apparatus in a semi-intact cell assay. Moreover, cytosol immunodepleted of mSEC13 could no longer support ER-Golgi transport. Transport could be restored in a dose-dependent manner by a cytosol fraction enriched in the high-molecular-weight mSEC13 complex but not by a fraction enriched in either monomeric mSEC13 or recombinant mSEC13. As a putative component of the mammalian COPII complex, mSEC13 showed partially overlapping but mostly different properties in terms of localization, membrane recruitment, and dynamics compared to that of beta-COP, a component of the COPI complex.

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