Identification and functional characterization of a novel human misshapen/Nck interacting kinase-related kinase, hMINK beta.
Misshapen/NIKs-related kinase (MINK) is a member of the germinal center family of kinases that are homologous to the yeast sterile 20 (Ste20) kinases and regulate a wide variety of cellular processes, including cell morphology, cytoskeletal rearrangement, and survival. Here, we present the cloning and functional characterization of a novel human Misshapen/NIKs-related kinase beta (hMINK beta) that encodes a polypeptide of 1312 amino acids. hMINK beta is ubiquitously expressed in most tissues with at least five alternatively spliced isoforms. Similar to Nck interacting kinase (NIK) and Traf2 and Nck-interacting kinase (TNIK), hMINK beta moderately activates c-Jun N-terminal kinase (JNK) and associates with Nck via the intermediate domain in the yeast two-hybrid system and in a glutathione S-transferase (GST) pull-down assay. Interestingly, overexpression of the kinase domain deleted and kinase-inactive mutants of hMINK beta in human fibrosarcoma HT1080 cells enhanced cell spreading, actin stress fiber formation, and adhesion to extracellular matrix, as well as decreased cell motility and cell invasion. Furthermore, these mutants also promoted cell-cell adhesion in human breast carcinoma MCF7 cells, evidenced with cell growth in clusters and increased membrane localization of beta-catenin, a multifunctional protein involved in E-cadherin-mediated cell adhesion. Finally, hMINK beta protein was found to colocalize with the Golgi apparatus, implicating that hMINK beta might exert its functions, at least in part, through the modulation of intracellular protein transport. Taken together, these results suggest that hMINK beta plays an important role in cytoskeleton reorganization, cell adhesion, and cell motility.
Hu
Yuanming
Y
Rigel Pharmaceuticals, Inc., South San Francisco, California 94080, USA.
Leo
Cindy
C
Yu
Simon
S
Huang
Betty C B
BC
Wang
Hank
H
Shen
Mary
M
Luo
Ying
Y
Daniel-Issakani
Sarkiz
S
Payan
Donald G
DG
Xu
Xiang
X
D048868:Adaptor Proteins, Signal Transducing; D017398:Alternative Splicing; D000818:Animals; D001665:Binding Sites; D015152:Blotting, Northern; D001943:Breast Neoplasms; D002448:Cell Adhesion; D002455:Cell Division; D002462:Cell Membrane / Q000737:chemistry; D002465:Cell Movement; D003001:Cloning, Molecular; D003598:Cytoskeletal Proteins / Q000032:analysis; D003599:Cytoskeleton / Q000502:physiology / Q000648:ultrastructure; D004789:Enzyme Activation; D005109:Extracellular Matrix; D005354:Fibrosarcoma; D005455:Fluorescent Antibody Technique; D017353:Gene Deletion; D015870:Gene Expression; D005982:Glutathione Transferase; D006056:Golgi Apparatus / Q000737:chemistry; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D048670:MAP Kinase Kinase 4; D051379:Mice; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis; D015513:Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D009928:Organ Specificity; D017354:Point Mutation; D017346:Protein Serine-Threonine Kinases / Q000032:analysis* / Q000235:genetics / Q000502:physiology*; D017385:Sequence Homology; D015534:Trans-Activators / Q000032:analysis; D014162:Transfection; D014407:Tumor Cells, Cultured; D020798:Two-Hybrid System Techniques; D051176:beta Catenin
D048868:Adaptor Proteins, Signal Transducing; C495270:CTNNB1 protein, human; C495267:CTNNB1 protein, mouse; D003598:Cytoskeletal Proteins; C078332:Nck protein; D015513:Oncogene Proteins; D015534:Trans-Activators; D051176:beta Catenin; D005982:Glutathione Transferase; C497353:MINK1 protein, human; D017346:Protein Serine-Threonine Kinases; D048031:JNK Mitogen-Activated Protein Kinases; D048670:MAP Kinase Kinase 4; D020929:Mitogen-Activated Protein Kinase Kinases