Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Subscribe

Logo for

J. Cell Biol. 153 (7): 1355-1368

Copyright © 2001 by the Rockefeller University Press.


Original Article

Tyrosine-Phosphorylated Extracellular Signal–Regulated Kinase Associates with the Golgi Complex during G2/M Phase of the Cell Cycle

Evidence for Regulation of Golgi Structure

Hyukjin Chaa, and Paul Shapiroa

a Department of Pharmaceutical Sciences, University of Maryland, School of Pharmacy, Baltimore, Maryland 21201
Department of Pharmaceutical Sciences, University of Maryland, School of Pharmacy, 20 N. Pine St., Baltimore, MD 21201.(410) 706-0346(410) 706-8522

pshapiro{at}rx.umaryland.edu

Abstract: Phosphorylation of the extracellular signal–regulated kinases (ERKs) on tyrosine and threonine residues within the TEY tripeptide motif induces ERK activation and targeting of substrates. Although it is recognized that phosphorylation of both residues is required for ERK activation, it is not known if a single phosphorylation of either residue regulates physiological functions. In light of recent evidence indicating that ERK proteins regulate substrate function in the absence of ERK enzymatic activity, we have begun to examine functional roles for partially phosphorylated forms of ERK. Using phosphorylation site–specific ERK antibodies and immunofluorescence, we demonstrate that ERK phosphorylated on the tyrosine residue (pY ERK) within the TEY activation sequence is found constitutively in the nucleus, and localizes to the Golgi complex of cells that are in late G2 or early mitosis of the cell cycle. As cells progress through metaphase and anaphase, pY ERK localization to Golgi vesicles is most evident around the mitotic spindle poles. During telophase, pY ERK associates with newly formed Golgi vesicles but is not found on there after cytokinesis and entry into G1. Increased ERK phosphorylation causes punctate distribution of several Golgi proteins, indicating disruption of the Golgi structure. This observation is reversible by overexpression of a tyrosine phosphorylation–defective ERK mutant, but not by a kinase-inactive ERK2 mutant that is tyrosine phosphorylated. These data provide the first evidence that pY ERK and not ERK kinase activity regulates Golgi structure and may be involved in mitotic Golgi fragmentation and reformation.

Key Words: mitosis • Golgi complex • tyrosine phosphorylation • MAP kinase • cell cycle



Abbreviations used in this paper: CENP-E, centromere protein E; ERK, extracellular signal–regulated kinase; HA, hemagglutinin; Mann II, mannosidase II; MAPK, mitogen-activated protein kinase; MEF, mouse embryonic fibroblast; MKK, MAPK kinase; pH3, phosphorylated histone H3; PKA, protein kinase A; PlK, polo-like kinase; pT ERK, threonine-phosphorylated ERK; pY ERK, tyrosine-phosphorylated ERK; pTpY ERK, dually phosphorylated ERK; Topo II{alpha}, topoisomerase II{alpha}.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Comparative analysis of Erk phosphorylation suggests a mixed strategy for measuring phospho-form distributions.
S. Prabakaran, R. A. Everley, I. Landrieu, J.-M. Wieruszeski, G. Lippens, H. Steen, and J. Gunawardena (2014)
Mol Syst Biol 7, 482
   Abstract »    Full Text »    PDF »
The ERK Cascade: Distinct Functions within Various Subcellular Organelles.
I. Wortzel and R. Seger (2011)
Genes & Cancer 2, 195-209
   Abstract »    Full Text »    PDF »
GRASP55 and GRASP65 play complementary and essential roles in Golgi cisternal stacking.
Y. Xiang and Y. Wang (2010)
J. Cell Biol. 188, 237-251
   Abstract »    Full Text »    PDF »
Specific phosphorylation and activation of ERK1c by MEK1b: a unique route in the ERK cascade.
Y. D. Shaul, G. Gibor, A. Plotnikov, and R. Seger (2009)
Genes & Dev. 23, 1779-1790
   Abstract »    Full Text »    PDF »
Plk3 Interacts with and Specifically Phosphorylates VRK1 in Ser342, a Downstream Target in a Pathway That Induces Golgi Fragmentation.
I. Lopez-Sanchez, M. Sanz-Garcia, and P. A. Lazo (2009)
Mol. Cell. Biol. 29, 1189-1201
   Abstract »    Full Text »    PDF »
Enzymatic Activity and Substrate Specificity of Mitogen-activated Protein Kinase p38{alpha} in Different Phosphorylation States.
Y.-Y. Zhang, Z.-Q. Mei, J.-W. Wu, and Z.-X. Wang (2008)
J. Biol. Chem. 283, 26591-26601
   Abstract »    Full Text »    PDF »
A Functional Role for p38 MAPK in Modulating Mitotic Transit in the Absence of Stress.
H. Cha, X. Wang, H. Li, and A. J. Fornace Jr. (2007)
J. Biol. Chem. 282, 22984-22992
   Abstract »    Full Text »    PDF »
Inhibition of the janus kinase family increases extracellular signal-regulated kinase 1/2 phosphorylation and causes endoreduplication..
G. Reiterer and A. Yen (2006)
Cancer Res. 66, 9083-9089
   Abstract »    Full Text »    PDF »
The physiology of membrane transport and endomembrane-based signalling.
M. Sallese, T. Pulvirenti, and A. Luini (2006)
EMBO J. 25, 2663-2673
   Abstract »    Full Text »    PDF »
ERK1c regulates Golgi fragmentation during mitosis..
Y. D. Shaul and R. Seger (2006)
J. Cell Biol. 172, 885-897
   Abstract »    Full Text »    PDF »
Oncogenic RAS Induces Accelerated Transition through G2/M and Promotes Defects in the G2 DNA Damage and Mitotic Spindle Checkpoints.
J. A. Knauf, B. Ouyang, E. S. Knudsen, K. Fukasawa, G. Babcock, and J. A. Fagin (2006)
J. Biol. Chem. 281, 3800-3809
   Abstract »    Full Text »    PDF »
Cdc2-mediated Inhibition of Epidermal Growth Factor Activation of the Extracellular Signal-regulated Kinase Pathway during Mitosis.
S. Dangi and P. Shapiro (2005)
J. Biol. Chem. 280, 24524-24531
   Abstract »    Full Text »    PDF »
Convergence of Cell Cycle Regulation and Growth Factor Signals on GRASP65.
S.-i. Yoshimura, K. Yoshioka, F. A. Barr, M. Lowe, K. Nakayama, S. Ohkuma, and N. Nakamura (2005)
J. Biol. Chem. 280, 23048-23056
   Abstract »    Full Text »    PDF »
Induction of Nitric Oxide Synthase in Anopheles stephensi by Plasmodium falciparum: Mechanism of Signaling and the Role of Parasite Glycosylphosphatidylinositols.
J. Lim, D. C. Gowda, G. Krishnegowda, and S. Luckhart (2005)
Infect. Immun. 73, 2778-2789
   Abstract »    Full Text »    PDF »
Protein Phosphatase 2A Activity Associated with Golgi Membranes during the G2/M Phase May Regulate Phosphorylation of ERK2.
C. N. Hancock, S. Dangi, and P. Shapiro (2005)
J. Biol. Chem. 280, 11590-11598
   Abstract »    Full Text »    PDF »
Molecular Determinants of Substrate Recognition in Hematopoietic Protein-tyrosine Phosphatase.
Z. Huang, B. Zhou, and Z.-Y. Zhang (2004)
J. Biol. Chem. 279, 52150-52159
   Abstract »    Full Text »    PDF »
Extracellular Signal-Regulated Kinase 1c (ERK1c), a Novel 42-Kilodalton ERK, Demonstrates Unique Modes of Regulation, Localization, and Function.
D. M. Aebersold, Y. D. Shaul, Y. Yung, N. Yarom, Z. Yao, T. Hanoch, and R. Seger (2004)
Mol. Cell. Biol. 24, 10000-10015
   Abstract »    Full Text »    PDF »
Cdc2 Kinase-dependent Disassembly of Endoplasmic Reticulum (ER) Exit Sites Inhibits ER-to-Golgi Vesicular Transport during Mitosis.
F. Kano, A. R. Tanaka, S. Yamauchi, H. Kondo, and M. Murata (2004)
Mol. Biol. Cell 15, 4289-4298
   Abstract »    Full Text »    PDF »
A Neuronal Isoform of Protein Kinase G Couples Mitogen-Activated Protein Kinase Nuclear Import to Axotomy-Induced Long-Term Hyperexcitability in Aplysia Sensory Neurons.
Y.-J. Sung, E. T. Walters, and R. T. Ambron (2004)
J. Neurosci. 24, 7583-7595
   Abstract »    Full Text »    PDF »
MAP kinase phosphatase-1 is induced in abnormal fibers in inclusion body myositis.
S. Nakano, A. Shinde, H. Ito, H. Ito, and H. Kusaka (2003)
Neurology 61, 322-326
   Abstract »    Full Text »    PDF »
RAF1-activated MEK1 is found on the Golgi apparatus in late prophase and is required for Golgi complex fragmentation in mitosis.
A. Colanzi, C. Sutterlin, and V. Malhotra (2003)
J. Cell Biol. 161, 27-32
   Abstract »    Full Text »    PDF »
The coiled-coil membrane protein golgin-84 is a novel rab effector required for Golgi ribbon formation.
A. Diao, D. Rahman, D. J.C. Pappin, J. Lucocq, and M. Lowe (2003)
J. Cell Biol. 160, 201-212
   Abstract »    Full Text »    PDF »
The Specificity of Extracellular Signal-regulated Kinase 2 Dephosphorylation by Protein Phosphatases.
B. Zhou, Z.-X. Wang, Y. Zhao, D. L. Brautigan, and Z.-Y. Zhang (2002)
J. Biol. Chem. 277, 31818-31825
   Abstract »    Full Text »    PDF »
The Activity of the Extracellular Signal-regulated Kinase 2 Is Regulated by Differential Phosphorylation in the Activation Loop.
B. Zhou and Z.-Y. Zhang (2002)
J. Biol. Chem. 277, 13889-13899
   Abstract »    Full Text »    PDF »
Identification of a C-terminal Region That Regulates Mitogen-activated Protein Kinase Kinase-1 Cytoplasmic Localization and ERK Activation.
H. Cha, E. K. Lee, and P. Shapiro (2001)
J. Biol. Chem. 276, 48494-48501
   Abstract »    Full Text »    PDF »
Signaling Pathways Regulating Golgi Structure and Function.
C. Preisinger and F. A. Barr (2001)
Sci. STKE 2001, pe38
   Abstract »    Full Text »    PDF »

To Advertise     Find Products


Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882