Evidence for Existence of a Nuclear Pore Complex-mediated, Cytosol-independent Pathway of Nuclear Translocation of ERK MAP Kinase in Permeabilized Cells

Matsubayashi, Yutaka; Fukuda, Makoto; Nishida, Eisuke

doi:10.1074/jbc.m106012200

Cited by 136 publications

(131 citation statements)

References 59 publications

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“…STRAD␣ (48 kDa) is similar in size to mitogen-activated protein kinase, which can diffuse passively through the pores, but there also exist at least 11 importins that might carry STRAD␣ into the nucleus (Khokhlatchev et al, 1998;Matsubayashi et al, 2001;Pemberton and Paschal, 2005). To examine the import mechanism of STRAD␣, we used recombinant His 6 -STRAD␣ labeled with OG-STRAD, GST-GFP-NLS (GGNLS), and His 6 -importin-␤-GFP.…”

Section: Strad␣ Enters the Nucleus By Passive Diffusionmentioning

confidence: 99%

STRADα Regulates LKB1 Localization by Blocking Access to Importin-α, and by Association with Crm1 and Exportin-7

Dorfman

Macara

2008

MBoC

109

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LKB1, a serine/threonine kinase, regulates cell polarity, metabolism, and cell growth. The activity and cellular distribution of LKB1 are determined by cofactors, STRAD␣ and MO25. STRAD␣ induces relocalization of LKB1 from the nucleus to the cytoplasm and stimulates its catalytic activity. MO25 stabilizes the STRAD␣/LKB1 interaction. We investigated the mechanism of nucleocytoplasmic transport of LKB1 in response to its cofactors. Although LKB1 is imported into the nucleus by importin-␣/␤, STRAD␣ and MO25 passively diffuse between the nucleus and the cytoplasm. STRAD␣ induces nucleocytoplasmic shuttling of LKB1. STRAD␣ facilitates nuclear export of LKB1 by serving as an adaptor between LKB1 and exportins CRM1 and exportin7. STRAD␣ inhibits import of LKB1 by competing with importin-␣ for binding to LKB1. MO25 stabilizes the LKB1-STRAD␣ complex but it does not facilitate its nucleocytoplasmic shuttling. Strikingly, the STRAD␤, isoform which differs from STRAD␣ in the N-and C-terminal domains that are responsible for interaction with export receptors, does not efficiently relocalize LKB1 from the nucleus to the cytoplasm. These results identify a multifactored mechanism to control LKB1 localization, and they suggest that the STRAD␤-LKB1 complex might possess unique functions in the nucleus. INTRODUCTIONPeutz-Jeghers cancer syndrome is caused by mutations in a gene encoding a serine/threonine kinase called LKB1 (Hemminki et al., 1998). This kinase is closely related to PAR-4, a protein required for polarization of the Caenorhabditis elegans zygote, and it has recently been implicated in the apical/basal polarization of mammalian epithelial cells (Baas et al., 2004). LKB1 is also involved in numerous cell signaling pathways that regulate cellular metabolism (Shaw et al., 2004Liang et al., 2007), cell growth, and responses to DNA damage (Wei et al., 2005;Setogawa et al., 2006;Zeng and Berger, 2006). LKB1 can phosphorylate and activate 13 protein kinases from the AMP-activated protein kinase family (Lizcano et al., 2004). However, the biological significance of this phosphorylation is still unclear for many of these kinases. The cellular localization and activity of LKB1 is controlled through its interaction with Ste20 Related Adaptor (STRAD) and an adapter protein, MO25 . STRAD, a 48-kDa protein, is a pseudokinase that consists of a STE20-like kinase domain but lacks several residues that are indespensible for catalytic activity. STRAD resembles most closely the STE20 homologues SPAK (Johnston et al., 2000) and ILPIP (Sanna et al., 2002), or PAP kinase (Nishigaki et al., 2003). Whereas SPAK is a true kinase, it was later shown that ILPIP/PAPK is a pseudokinase, similarly to STRAD and that it also interacts with and activates LKB1. Therefore, it was termed STRAD␤, whereas the original STRAD became known as STRAD␣ . The complex of LKB1 and STRAD␣ or -␤ is stabilized by MO25, a 40-kDa protein composed of ␣-helical repeats that are distantly related to the armadillo repeat domain (Milburn et al., 2004). MO25 binds STRAD ...

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Section: Strad␣ Enters the Nucleus By Passive Diffusionmentioning

confidence: 99%

STRADα Regulates LKB1 Localization by Blocking Access to Importin-α, and by Association with Crm1 and Exportin-7

Dorfman

Macara

2008

MBoC

109

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“…In particular, ERK, which is activated upon phosphorylation by dual specificity MEK1 and MEK2 (13), can have biological effects by phosphorylating membrane or cytoskeletal proteins (14). Moreover, when phosphorylated ERK (pERK) translocates to the nucleus (15,16), it can result in activation of transcription factors, including activator protein-1 (AP-1) (17).…”

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confidence: 99%

Helicobacter pylori Induces ERK-dependent Formation of a Phospho-c-Fos·c-Jun Activator Protein-1 Complex That Causes Apoptosis in Macrophages

Asim

Chaturvedi

Hoge

et al. 2010

Journal of Biological Chemistry

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Macrophages are essential components of innate immunity, and apoptosis of these cells impairs mucosal defense to microbes. Helicobacter pylori is a gastric pathogen that infects half of the world population and causes peptic ulcer disease and gastric cancer. The host inflammatory response fails to eradicate the organism. We have reported that H. pylori induces apoptosis of macrophages by generation of polyamines from ornithine decarboxylase (

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“…Overexpression studies have shown that ERK2 export can be facilitated by MEK1, which has a nuclear export sequence (NES) and requires the NES-binding protein CRM1 for movement out of the nucleus (16,17). Nuclear entry of ERK2 has been described recently as a facilitated mechanism in which ERK2 moves through the NPC by binding directly to nucleoporins (18,19). This process, which has been described for at least two other signaling proteins (20,21), does not rely on the classical, karyopherin-mediated import mechanism.…”

mentioning

confidence: 99%

The Death Effector Domain Protein PEA-15 Prevents Nuclear Entry of ERK2 by Inhibiting Required Interactions

Whitehurst

Robinson

Moore

et al. 2004

Journal of Biological Chemistry

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ERK2 nuclear-cytoplasmic distribution is regulated in response to hormones and cellular state without the requirement for karyopherin-mediated nuclear import. One proposed mechanism for the movement of ERK2 into the nucleus is through a direct interaction between ERK2 and nucleoporins present in the nuclear pore complex. Previous reports have attributed regulation of ERK2 localization to proteins that activate or deactivate ERK2, such as the mitogen-activated protein (MAP) kinase kinase MEK1 and MAP kinase phosphatases. Recently, a small non-catalytic protein, PEA-15, has also been demonstrated to promote a cytoplasmic ERK2 localization. We found that the MAP kinase insert in ERK2 is required for its interaction with PEA-15. Consistent with its recognition of the MAP kinase insert, PEA-15 blocked activation of ERK2 by MEK1, which also requires the MAP kinase insert to interact productively with ERK2. To determine how PEA-15 influences the localization of ERK2, we used a permeabilized cell system to examine the effect of PEA-15 on the localization of ERK2 and mutants that have lost the ability to bind PEA-15. Wild type ERK2 was unable to enter the nucleus in the presence of an excess of PEA-15; however, ERK2 lacking the MAP kinase insert largely retained the ability to enter the nucleus. Binding assays demonstrated that PEA-15 interfered with the ability of ERK2 to bind to nucleoporins. These results suggest that PEA-15 sequesters ERK2 in the cytoplasm at least in part by interfering with its ability to interact with nucleoporins, presenting a potential paradigm for regulation of ERK2 localization.The mitogen-activated protein (MAP) 1 kinase, ERK2, plays a critical role in promoting cellular changes in response to both mitogenic and non-mitogenic stimuli. ERK2 is the multifunctional kinase in a kinase cascade that is activated by various stimuli acting through tyrosine kinase receptors, G proteincoupled receptors, and others (1, 2). Activation of this cascade results in the dual phosphorylation of ERK2 and the consequent phosphorylation of target kinases, transcription factors, and other proteins throughout the cell. The effectors of ERK2 are localized in both the cytoplasm and the nucleus, making its subcellular localization important for its ability to induce cellular changes (3, 4).Regulation of ERK2 localization has been characterized mostly by overexpression and/or by using antibodies to N-and C-terminal epitopes (5, 6); these epitopes are not readily accessible in ERK2 that is associated with microtubules, which constitutes a large portion of the cytoplasmic protein (7). ERK2 and its upstream activator the MAP/ERK kinase MEK1 (also known as MKK1) interact stably through multiple sites with an affinity in the micromolar range (8 -14). Overexpressed ERK2 accumulates in the nucleus in a stimulus-independent manner. This suggests that cytosolic binding is important in determining the distribution of ERK2 in cells. Coexpression with MEK1 shifts the distribution in favor of the cytoplasm (13). From these studies,...

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Evidence for Existence of a Nuclear Pore Complex-mediated, Cytosol-independent Pathway of Nuclear Translocation of ERK MAP Kinase in Permeabilized Cells

Cited by 136 publications

References 59 publications

STRADα Regulates LKB1 Localization by Blocking Access to Importin-α, and by Association with Crm1 and Exportin-7

STRADα Regulates LKB1 Localization by Blocking Access to Importin-α, and by Association with Crm1 and Exportin-7

Helicobacter pylori Induces ERK-dependent Formation of a Phospho-c-Fos·c-Jun Activator Protein-1 Complex That Causes Apoptosis in Macrophages

The Death Effector Domain Protein PEA-15 Prevents Nuclear Entry of ERK2 by Inhibiting Required Interactions

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