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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


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}.

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