Tags

Type your tag names separated by a space and hit enter

Biochemical and biological functions of the N-terminal, noncatalytic domain of extracellular signal-regulated kinase 2.
Mol Cell Biol. 2001 Jan; 21(1):249-59.MC

Abstract

Extracellular signal-regulated kinase 1 (ERK1) and ERK2 are important components in signal transduction pathways involved in many cellular processes, including cell differentiation and proliferation. These proteins consist of a central kinase domain flanked by short N- and C-terminal noncatalytic domains. While the regulation of ERK2 by sequences within the kinase domain has been extensively studied, little is known about the small regions outside of the kinase domain. We performed mutational analysis on the N-terminal, noncatalytic domain of ERK2 in an attempt to determine its role in ERK2 function and regulation. Deleting or mutating amino acids 19 to 25 (ERK2-Delta19-25) created an ERK2 molecule that could be phosphorylated in response to growth factor and serum stimulation in a MEK (mitogen-activated protein kinase kinase or ERK kinase)-dependent manner but had little kinase activity and was unable to bind to MEK in vivo. Since MEK acts as a cytoplasmic anchor for the ERKs, the lack of a MEK interaction resulted in the aberrant nuclear localization of ERK2-Delta19-25 mutants in serum-starved cells. Assaying these mutants for their ability to affect ERK signaling revealed that ERK2-Delta19-25 mutants acted in a dominant-negative manner to inhibit transcriptional signaling through endogenous ERKs to an Elk1-responsive promoter in transfected COS-1 cells. However, ERK2-Delta19-25 had no effect on the phosphorylation of RSK2, an ERK2 cytoplasmic substrate, whereas a nonactivatable ERK (T183A) that retained these sequences could inhibit RSK2 phosphorylation. These results suggest that the N-terminal domain of ERK2 profoundly affects ERK2 localization, MEK binding, kinase activity, and signaling and identify a novel dominant-negative mutant of ERK2 that can dissociate at least some transcriptional responses from cytoplasmic responses.

Authors+Show Affiliations

Department of Microbiology and Cancer Center, School of Medicine, University of Virginia, Charlottesville, Virginia 22908, USA.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

11113199

Citation

Eblen, S T., et al. "Biochemical and Biological Functions of the N-terminal, Noncatalytic Domain of Extracellular Signal-regulated Kinase 2." Molecular and Cellular Biology, vol. 21, no. 1, 2001, pp. 249-59.
Eblen ST, Catling AD, Assanah MC, et al. Biochemical and biological functions of the N-terminal, noncatalytic domain of extracellular signal-regulated kinase 2. Mol Cell Biol. 2001;21(1):249-59.
Eblen, S. T., Catling, A. D., Assanah, M. C., & Weber, M. J. (2001). Biochemical and biological functions of the N-terminal, noncatalytic domain of extracellular signal-regulated kinase 2. Molecular and Cellular Biology, 21(1), 249-59.
Eblen ST, et al. Biochemical and Biological Functions of the N-terminal, Noncatalytic Domain of Extracellular Signal-regulated Kinase 2. Mol Cell Biol. 2001;21(1):249-59. PubMed PMID: 11113199.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Biochemical and biological functions of the N-terminal, noncatalytic domain of extracellular signal-regulated kinase 2. AU - Eblen,S T, AU - Catling,A D, AU - Assanah,M C, AU - Weber,M J, PY - 2000/12/13/pubmed PY - 2001/2/28/medline PY - 2000/12/13/entrez SP - 249 EP - 59 JF - Molecular and cellular biology JO - Mol. Cell. Biol. VL - 21 IS - 1 N2 - Extracellular signal-regulated kinase 1 (ERK1) and ERK2 are important components in signal transduction pathways involved in many cellular processes, including cell differentiation and proliferation. These proteins consist of a central kinase domain flanked by short N- and C-terminal noncatalytic domains. While the regulation of ERK2 by sequences within the kinase domain has been extensively studied, little is known about the small regions outside of the kinase domain. We performed mutational analysis on the N-terminal, noncatalytic domain of ERK2 in an attempt to determine its role in ERK2 function and regulation. Deleting or mutating amino acids 19 to 25 (ERK2-Delta19-25) created an ERK2 molecule that could be phosphorylated in response to growth factor and serum stimulation in a MEK (mitogen-activated protein kinase kinase or ERK kinase)-dependent manner but had little kinase activity and was unable to bind to MEK in vivo. Since MEK acts as a cytoplasmic anchor for the ERKs, the lack of a MEK interaction resulted in the aberrant nuclear localization of ERK2-Delta19-25 mutants in serum-starved cells. Assaying these mutants for their ability to affect ERK signaling revealed that ERK2-Delta19-25 mutants acted in a dominant-negative manner to inhibit transcriptional signaling through endogenous ERKs to an Elk1-responsive promoter in transfected COS-1 cells. However, ERK2-Delta19-25 had no effect on the phosphorylation of RSK2, an ERK2 cytoplasmic substrate, whereas a nonactivatable ERK (T183A) that retained these sequences could inhibit RSK2 phosphorylation. These results suggest that the N-terminal domain of ERK2 profoundly affects ERK2 localization, MEK binding, kinase activity, and signaling and identify a novel dominant-negative mutant of ERK2 that can dissociate at least some transcriptional responses from cytoplasmic responses. SN - 0270-7306 UR - https://www.unboundmedicine.com/medline/citation/11113199/Biochemical_and_biological_functions_of_the_N_terminal_noncatalytic_domain_of_extracellular_signal_regulated_kinase_2_ L2 - http://mcb.asm.org/cgi/pmidlookup?view=long&pmid=11113199 DB - PRIME DP - Unbound Medicine ER -