Endoplasmic reticulum export sites and golgi bodies behave as single mobile secretory units in plant cells

Luis L P DaSilva, Erik L. Snapp, Jürgen Denecke, Jennifer Lippincott-Schwarb, Chris Hawes, Federica Brandizzi

Research output: Contribution to journalArticle

211 Citations (Scopus)

Abstract

In contrast with animals, plant cells contain multiple mobile Golgi stacks distributed over the entire cytoplasm. However, the distribution and dynamics of protein export sites on the plant endoplasmic reticulum (ER) surface have yet to be characterized. A widely accepted model for ER-to-Golgi transport is based on the sequential action of COPII and COPI coat complexes. The COPII complex assembles by the ordered recruitment of cytosolic components on the ER membrane. Here, we have visualized two early components of the COPII machinery, the small GTPase Sar1p and its GTP exchanging factor Sec12p in live tobacco (Nicotiana tabacum) leaf epidermal cells. By in vivo confocal laser scanning microscopy and fluorescence recovery after photobleaching experiments, we show that Sar1p cycles on mobile punctate structures that track with the Golgi bodies in close proximity but contain regions that are physically separated from the Golgi bodies. By contrast, Sec12p is uniformly distributed along the ER network and does not accumulate in these structures, consistent with the fact that Sec12p does not become part of a COPII vesicle. We propose that punctate accumulation of Sar1p represents ER export sites (ERES). The sites may represent a combination of Sar1p-coated ER membranes, nascent COPII membranes, and COPII vectors in transit, which have yet to lose their coats. ERES can be induced by overproducing Golgi membrane proteins but not soluble bulk-flow cargos. Few punctate Sar1p loci were observed that are independent of Golgi bodies, and these may be nascent ERES. The vast majority of ERES form secretory units that move along the surface of the ER together with the Golgi bodies, but movement does not influence the rate of cargo transport between these two organelles. Moreover, we could demonstrate using the drug brefeldin A that formation of ERES is strictly dependent on a functional retrograde transport route from the Golgi apparatus.

Original languageEnglish (US)
Pages (from-to)1753-1771
Number of pages19
JournalPlant Cell
Volume16
Issue number7
DOIs
StatePublished - Jul 2004
Externally publishedYes

Fingerprint

Plant Cells
Golgi apparatus
Endoplasmic Reticulum
endoplasmic reticulum
Membranes
Coat Protein Complex I
Brefeldin A
Photobleaching
Monomeric GTP-Binding Proteins
Tobacco
cells
Guanosine Triphosphate
Machinery
Microscopic examination
Membrane Proteins
Animals
Fluorescence
Scanning
Recovery
Fluorescence Recovery After Photobleaching

ASJC Scopus subject areas

  • Plant Science
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

DaSilva, L. L. P., Snapp, E. L., Denecke, J., Lippincott-Schwarb, J., Hawes, C., & Brandizzi, F. (2004). Endoplasmic reticulum export sites and golgi bodies behave as single mobile secretory units in plant cells. Plant Cell, 16(7), 1753-1771. https://doi.org/10.1105/tpc.022673

Endoplasmic reticulum export sites and golgi bodies behave as single mobile secretory units in plant cells. / DaSilva, Luis L P; Snapp, Erik L.; Denecke, Jürgen; Lippincott-Schwarb, Jennifer; Hawes, Chris; Brandizzi, Federica.

In: Plant Cell, Vol. 16, No. 7, 07.2004, p. 1753-1771.

Research output: Contribution to journalArticle

DaSilva, LLP, Snapp, EL, Denecke, J, Lippincott-Schwarb, J, Hawes, C & Brandizzi, F 2004, 'Endoplasmic reticulum export sites and golgi bodies behave as single mobile secretory units in plant cells', Plant Cell, vol. 16, no. 7, pp. 1753-1771. https://doi.org/10.1105/tpc.022673
DaSilva LLP, Snapp EL, Denecke J, Lippincott-Schwarb J, Hawes C, Brandizzi F. Endoplasmic reticulum export sites and golgi bodies behave as single mobile secretory units in plant cells. Plant Cell. 2004 Jul;16(7):1753-1771. https://doi.org/10.1105/tpc.022673
DaSilva, Luis L P ; Snapp, Erik L. ; Denecke, Jürgen ; Lippincott-Schwarb, Jennifer ; Hawes, Chris ; Brandizzi, Federica. / Endoplasmic reticulum export sites and golgi bodies behave as single mobile secretory units in plant cells. In: Plant Cell. 2004 ; Vol. 16, No. 7. pp. 1753-1771.
@article{7ada2573ece5400e8b68272aa5397ea3,
title = "Endoplasmic reticulum export sites and golgi bodies behave as single mobile secretory units in plant cells",
abstract = "In contrast with animals, plant cells contain multiple mobile Golgi stacks distributed over the entire cytoplasm. However, the distribution and dynamics of protein export sites on the plant endoplasmic reticulum (ER) surface have yet to be characterized. A widely accepted model for ER-to-Golgi transport is based on the sequential action of COPII and COPI coat complexes. The COPII complex assembles by the ordered recruitment of cytosolic components on the ER membrane. Here, we have visualized two early components of the COPII machinery, the small GTPase Sar1p and its GTP exchanging factor Sec12p in live tobacco (Nicotiana tabacum) leaf epidermal cells. By in vivo confocal laser scanning microscopy and fluorescence recovery after photobleaching experiments, we show that Sar1p cycles on mobile punctate structures that track with the Golgi bodies in close proximity but contain regions that are physically separated from the Golgi bodies. By contrast, Sec12p is uniformly distributed along the ER network and does not accumulate in these structures, consistent with the fact that Sec12p does not become part of a COPII vesicle. We propose that punctate accumulation of Sar1p represents ER export sites (ERES). The sites may represent a combination of Sar1p-coated ER membranes, nascent COPII membranes, and COPII vectors in transit, which have yet to lose their coats. ERES can be induced by overproducing Golgi membrane proteins but not soluble bulk-flow cargos. Few punctate Sar1p loci were observed that are independent of Golgi bodies, and these may be nascent ERES. The vast majority of ERES form secretory units that move along the surface of the ER together with the Golgi bodies, but movement does not influence the rate of cargo transport between these two organelles. Moreover, we could demonstrate using the drug brefeldin A that formation of ERES is strictly dependent on a functional retrograde transport route from the Golgi apparatus.",
author = "DaSilva, {Luis L P} and Snapp, {Erik L.} and J{\"u}rgen Denecke and Jennifer Lippincott-Schwarb and Chris Hawes and Federica Brandizzi",
year = "2004",
month = "7",
doi = "10.1105/tpc.022673",
language = "English (US)",
volume = "16",
pages = "1753--1771",
journal = "Plant Cell",
issn = "1040-4651",
publisher = "American Society of Plant Biologists",
number = "7",

}

TY - JOUR

T1 - Endoplasmic reticulum export sites and golgi bodies behave as single mobile secretory units in plant cells

AU - DaSilva, Luis L P

AU - Snapp, Erik L.

AU - Denecke, Jürgen

AU - Lippincott-Schwarb, Jennifer

AU - Hawes, Chris

AU - Brandizzi, Federica

PY - 2004/7

Y1 - 2004/7

N2 - In contrast with animals, plant cells contain multiple mobile Golgi stacks distributed over the entire cytoplasm. However, the distribution and dynamics of protein export sites on the plant endoplasmic reticulum (ER) surface have yet to be characterized. A widely accepted model for ER-to-Golgi transport is based on the sequential action of COPII and COPI coat complexes. The COPII complex assembles by the ordered recruitment of cytosolic components on the ER membrane. Here, we have visualized two early components of the COPII machinery, the small GTPase Sar1p and its GTP exchanging factor Sec12p in live tobacco (Nicotiana tabacum) leaf epidermal cells. By in vivo confocal laser scanning microscopy and fluorescence recovery after photobleaching experiments, we show that Sar1p cycles on mobile punctate structures that track with the Golgi bodies in close proximity but contain regions that are physically separated from the Golgi bodies. By contrast, Sec12p is uniformly distributed along the ER network and does not accumulate in these structures, consistent with the fact that Sec12p does not become part of a COPII vesicle. We propose that punctate accumulation of Sar1p represents ER export sites (ERES). The sites may represent a combination of Sar1p-coated ER membranes, nascent COPII membranes, and COPII vectors in transit, which have yet to lose their coats. ERES can be induced by overproducing Golgi membrane proteins but not soluble bulk-flow cargos. Few punctate Sar1p loci were observed that are independent of Golgi bodies, and these may be nascent ERES. The vast majority of ERES form secretory units that move along the surface of the ER together with the Golgi bodies, but movement does not influence the rate of cargo transport between these two organelles. Moreover, we could demonstrate using the drug brefeldin A that formation of ERES is strictly dependent on a functional retrograde transport route from the Golgi apparatus.

AB - In contrast with animals, plant cells contain multiple mobile Golgi stacks distributed over the entire cytoplasm. However, the distribution and dynamics of protein export sites on the plant endoplasmic reticulum (ER) surface have yet to be characterized. A widely accepted model for ER-to-Golgi transport is based on the sequential action of COPII and COPI coat complexes. The COPII complex assembles by the ordered recruitment of cytosolic components on the ER membrane. Here, we have visualized two early components of the COPII machinery, the small GTPase Sar1p and its GTP exchanging factor Sec12p in live tobacco (Nicotiana tabacum) leaf epidermal cells. By in vivo confocal laser scanning microscopy and fluorescence recovery after photobleaching experiments, we show that Sar1p cycles on mobile punctate structures that track with the Golgi bodies in close proximity but contain regions that are physically separated from the Golgi bodies. By contrast, Sec12p is uniformly distributed along the ER network and does not accumulate in these structures, consistent with the fact that Sec12p does not become part of a COPII vesicle. We propose that punctate accumulation of Sar1p represents ER export sites (ERES). The sites may represent a combination of Sar1p-coated ER membranes, nascent COPII membranes, and COPII vectors in transit, which have yet to lose their coats. ERES can be induced by overproducing Golgi membrane proteins but not soluble bulk-flow cargos. Few punctate Sar1p loci were observed that are independent of Golgi bodies, and these may be nascent ERES. The vast majority of ERES form secretory units that move along the surface of the ER together with the Golgi bodies, but movement does not influence the rate of cargo transport between these two organelles. Moreover, we could demonstrate using the drug brefeldin A that formation of ERES is strictly dependent on a functional retrograde transport route from the Golgi apparatus.

UR - http://www.scopus.com/inward/record.url?scp=3142706602&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=3142706602&partnerID=8YFLogxK

U2 - 10.1105/tpc.022673

DO - 10.1105/tpc.022673

M3 - Article

VL - 16

SP - 1753

EP - 1771

JO - Plant Cell

JF - Plant Cell

SN - 1040-4651

IS - 7

ER -