Advanced

Identification of a C-terminal region that is required for the nuclear translocation of ERK2 by passive diffusion

Shibayama, Sotaro LU ; Shibata-Seita, Ryoko; Miura, Kenji; Kirino, Yutaka and Takishima, Kunio (2002) In Journal of Biological Chemistry 277(40). p.37777-37782
Abstract

Extracellular signal-regulated kinase 2 (ERK2) is located in the cytoplasm of resting cells and translocates into the nucleus upon extracellular stimuli by active transport of a dimer. Passive transport of an ERK2 monomer through the nuclear pore is also reported to coexist. We attempted to characterize the cytoplasmic retention and nuclear translocation of fusion proteins between deletion and site-directed mutants of ERK2 and green fluorescent protein (GFP). The overexpressed ERK2-GFP fusion protein is usually localized to both the cytoplasm and the nucleus unless a cytoplasmic anchoring protein is coexpressed. Deletion of 45 residues, but not 43 residues, from the C terminus of ERK2 prevented the nuclear distribution of the ERK2-GFP... (More)

Extracellular signal-regulated kinase 2 (ERK2) is located in the cytoplasm of resting cells and translocates into the nucleus upon extracellular stimuli by active transport of a dimer. Passive transport of an ERK2 monomer through the nuclear pore is also reported to coexist. We attempted to characterize the cytoplasmic retention and nuclear translocation of fusion proteins between deletion and site-directed mutants of ERK2 and green fluorescent protein (GFP). The overexpressed ERK2-GFP fusion protein is usually localized to both the cytoplasm and the nucleus unless a cytoplasmic anchoring protein is coexpressed. Deletion of 45 residues, but not 43 residues, from the C terminus of ERK2 prevented the nuclear distribution of the ERK2-GFP fusion protein. Substitution of a part of residues 299-313 to alanine residues also prevented the nuclear distribution of the ERK2-GFP fusion protein without abrogation of its nuclear active transport. These observations may indicate that the passive diffusion of ERK2 into the nucleus is not simple diffusion but includes a specific interaction process between residues 299-313 and the nuclear pore complex and that this interaction is not required for the active transport. We also showed that substitution of Tyr314 to alanine residue abrogated the cytoplasmic retention of the ERK2-GFP fusion protein by PTP-SL but not by MEK1.

(Less)
Please use this url to cite or link to this publication:
author
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biological Chemistry
volume
277
issue
40
pages
6 pages
publisher
ASBMB
external identifiers
  • scopus:0037020245
ISSN
0021-9258
DOI
10.1074/jbc.M206163200
language
English
LU publication?
no
id
1ef9abd1-1083-41ad-a0b6-cd4a639b3477
date added to LUP
2017-04-19 16:42:46
date last changed
2017-04-25 16:07:15
@article{1ef9abd1-1083-41ad-a0b6-cd4a639b3477,
  abstract     = {<p>Extracellular signal-regulated kinase 2 (ERK2) is located in the cytoplasm of resting cells and translocates into the nucleus upon extracellular stimuli by active transport of a dimer. Passive transport of an ERK2 monomer through the nuclear pore is also reported to coexist. We attempted to characterize the cytoplasmic retention and nuclear translocation of fusion proteins between deletion and site-directed mutants of ERK2 and green fluorescent protein (GFP). The overexpressed ERK2-GFP fusion protein is usually localized to both the cytoplasm and the nucleus unless a cytoplasmic anchoring protein is coexpressed. Deletion of 45 residues, but not 43 residues, from the C terminus of ERK2 prevented the nuclear distribution of the ERK2-GFP fusion protein. Substitution of a part of residues 299-313 to alanine residues also prevented the nuclear distribution of the ERK2-GFP fusion protein without abrogation of its nuclear active transport. These observations may indicate that the passive diffusion of ERK2 into the nucleus is not simple diffusion but includes a specific interaction process between residues 299-313 and the nuclear pore complex and that this interaction is not required for the active transport. We also showed that substitution of Tyr<sup>314</sup> to alanine residue abrogated the cytoplasmic retention of the ERK2-GFP fusion protein by PTP-SL but not by MEK1.</p>},
  author       = {Shibayama, Sotaro and Shibata-Seita, Ryoko and Miura, Kenji and Kirino, Yutaka and Takishima, Kunio},
  issn         = {0021-9258},
  language     = {eng},
  month        = {10},
  number       = {40},
  pages        = {37777--37782},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Identification of a C-terminal region that is required for the nuclear translocation of ERK2 by passive diffusion},
  url          = {http://dx.doi.org/10.1074/jbc.M206163200},
  volume       = {277},
  year         = {2002},
}