An association between NUAK2 and MRIP reveals a novel mechanism for regulation of actin stress fibers.
Actin stress fiber assembly and contractility in nonmuscle motile cells requires phosphorylation of myosin regulatory light chain (MLC). Dephosphorylation and disassembly are mediated by MLC phosphatase, which is targeted to actin fibers by the association of its regulatory subunit MYPT1 with myosin phosphatase Rho-interacting protein (MRIP). In the present study, we identify the kinase NUAK2 as a second protein targeted by MRIP to actin fibers. Association of NUAK2 with MRIP increases MLC phosphorylation and promotes formation of stress fibers. This activity does not require the kinase activity of NUAK2 but is dependent on both MRIP and MYPT1, indicating that the NUAK2-MRIP association inhibits fiber disassembly and MYPT1-mediated MLC dephosphorylation. NUAK2 levels are strongly induced by stimuli increasing actomyosin fiber formation, and NUAK2 is required for fiber maintenance in exponentially growing cells, implicating NUAK2 in a positive-feedback loop regulating actin stress fibers independently of the MLC kinase Rho-associated protein kinase (ROCK). The identified MRIP-NUAK2 association reveals a novel mechanism for the maintenance of actin stress fibers through counteracting MYPT1 and, together with recent results, implicates the NUAK proteins as important regulators of the MLC phosphatase acting in both a kinase-dependent and kinase-independent manner.
Institute of Biotechnology and Genome-Scale Biology Program, University of Helsinki, PO Box 56 Viikinkaari 9, 00014 University of Helsinki, Finland. tea.vallenius@helsinki.fi
Tea
T
Vallenius
Kari
K
Vaahtomeri
Bianca
B
Kovac
Ana-Maria
AM
Osiceanu
Martta
M
Viljanen
Tomi P
TP
Mäkelä
D000199:Actins / Q000378:metabolism*; D045744:Cell Line, Tumor; D019204:GTP-Binding Proteins / Q000378:metabolism*; D020690:GTPase-Activating Proteins / Q000378:metabolism*; D006801:Humans; D009119:Muscle Contraction; D018994:Myosin Light Chains / Q000378:metabolism; D009219:Myosin-Light-Chain Kinase / Q000378:metabolism; D043263:Myosin-Light-Chain Phosphatase / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D022502:Stress Fibers / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D054460:rho-Associated Kinases / Q000378:metabolism
D000199:Actins; D020690:GTPase-Activating Proteins; D018994:Myosin Light Chains; C486061:NUAK2 protein, human; D017346:Protein Serine-Threonine Kinases; D054460:rho-Associated Kinases; D009219:Myosin-Light-Chain Kinase; D043263:Myosin-Light-Chain Phosphatase; C490372:PPP1R12A protein, human; C472718:AGAP3 protein, human; D019204:GTP-Binding Proteins