PMID- 10037701
OWN - NLM
STAT- MEDLINE
DCOM- 19990330
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 10
DP  - 1999 Mar 5
TI  - Nuclear localization of mitogen-activated protein kinase kinase 1 (MKK1) is
      promoted by serum stimulation and G2-M progression. Requirement for
      phosphorylation at the activation lip and signaling downstream of MKK.
PG  - 6168-74
AB  - Stimulation of mammalian cells results in subcellular relocalization of Ras
      pathway enzymes, in which extracellular signal-regulated protein kinases rapidly 
      translocate to nuclei. In this study, we define conditions for nuclear
      localization of mitogen-activated protein kinase kinase 1 (MKK1) by examining
      effects of perturbing the nuclear export signal (NES), the regulatory
      phosphorylation sites Ser218 and Ser222, and a regulatory domain at the N
      terminus. After disrupting the NES (Delta32-37), nuclear uptake of MKK was
      enhanced when quiescent cells were activated with serum-phorbol 12-myristate
      13-acetate or BXB-Raf-1 cotransfection. Uptake was enhanced by mutation of Ser218
      and Ser222 to Glu and Asp, respectively, and blocked by mutation of these
      residues to Ala, although mutation of Lys97 to Met, which renders MKK
      catalytically inactive, did not interfere with uptake. Therefore, nuclear uptake 
      of MKK requires incorporation of phosphate or negatively charged residues at the 
      activation lip but not enzyme activity. On the other hand, uptake of an active
      MKK mutant with disrupted NES (Delta32-51) was elevated in quiescent as well as
      stimulated cells, and pretreatment of cells with the MKK inhibitor 1,4-diamino-2,
      3-dicyano-1,4-bis[2-aminophenylthio]butadiene blocked nuclear uptake. Thus,
      signaling downstream of MKK is also necessary for translocation. Finally, wild
      type MKK containing an intact NES translocates to nuclei during mitosis before
      envelope breakdown. Comparison of mutants with Ser to Glu and Asp or Ala
      substitutions indicates that Ser phosphorylation is also required for mitotic
      nuclear uptake of MKK.
FAU - Tolwinski, N S
AU  - Tolwinski NS
AD  - Department of Chemistry and Biochemistry, University of Colorado, Boulder,
      Colorado 80309, USA.
FAU - Shapiro, P S
AU  - Shapiro PS
FAU - Goueli, S
AU  - Goueli S
FAU - Ahn, N G
AU  - Ahn NG
LA  - eng
GR  - F32 GM18151/GM/NIGMS NIH HHS/United States
GR  - R01-GM48521/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - EC 2.7.10.1 (Protein-Tyrosine Kinases)
RN  - EC 2.7.11.1 (Protein-Serine-Threonine Kinases)
RN  - EC 2.7.12.2 (MAP Kinase Kinase 1)
RN  - EC 2.7.12.2 (Map2k1 protein, mouse)
RN  - EC 2.7.12.2 (Mitogen-Activated Protein Kinase Kinases)
SB  - IM
MH  - 3T3 Cells
MH  - Animals
MH  - Biological Transport
MH  - Cell Nucleus/*enzymology/ultrastructure
MH  - G2 Phase
MH  - MAP Kinase Kinase 1
MH  - Mice
MH  - *Mitogen-Activated Protein Kinase Kinases
MH  - Mitosis
MH  - Phosphorylation
MH  - Protein-Serine-Threonine Kinases/*metabolism
MH  - Protein-Tyrosine Kinases/*metabolism
MH  - *Signal Transduction
EDAT- 1999/02/26 00:00
MHDA- 1999/02/26 00:01
CRDT- 1999/02/26 00:00
PHST- 1999/02/26 00:00 [pubmed]
PHST- 1999/02/26 00:01 [medline]
PHST- 1999/02/26 00:00 [entrez]
AID - 10.1074/jbc.274.10.6168 [doi]
AID - S0021-9258(19)87570-1 [pii]
PST - ppublish
SO  - J Biol Chem. 1999 Mar 5;274(10):6168-74. doi: 10.1074/jbc.274.10.6168.