Molecular cloning and characterization of a novel member of the MAP kinase superfamily

Miyata, Yoshihiko; Akashi, Makoto; Nishida, Eisuke

doi:10.1046/j.1365-2443.1999.00261.x

Cited by 78 publications

(85 citation statements)

References 39 publications

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“…Interestingly, we could not find an obvious homologue in Plasmodium, which has cilia, but the cilia are assembled in the cytoplasm in a process that does not require IFT (42). Mammals have three putative DYF-5 homologues that form a small subfamily of MAP kinases, the MAK kinases (22)(23)(24)(25). The functions of these MAK, MOK, and ICK/MRK proteins remain to be determined.…”

Section: Discussionmentioning

confidence: 86%

“…dyf-5 encodes a protein of 471 aa, which shows extensive homology to three mammalian proteins that form a small subfamily of MAP kinases. This subfamily consists of MAK (male germ cell-associated kinase) (22), ICK or MRK (intestinal cell kinase and MAKrelated kinase, respectively) (23,24), and MOK (MAPK/MAK/ MRK overlapping kinase) (25), with highly conserved N-terminal catalytic domains and more divergent C-terminal noncatalytic domains. Although the functions of these kinases in mammals are not known, studies in other organisms suggest a function in the cilia.…”

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confidence: 99%

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Mutation of the MAP kinase DYF-5 affects docking and undocking of kinesin-2 motors and reduces their speed in the cilia of Caenorhabditis elegans

Burghoorn¹,

Dekkers²,

Rademakers³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

116

178

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In the cilia of the nematode Caenorhabditis elegans, anterograde intraflagellar transport (IFT) is mediated by two kinesin-2 complexes, kinesin II and OSM-3 kinesin. These complexes function together in the cilia middle segments, whereas OSM-3 alone mediates transport in the distal segments. Not much is known about the mechanisms that compartmentalize the kinesin-2 complexes or how transport by both kinesins is coordinated. Here, we identify DYF-5, a conserved MAP kinase that plays a role in these processes. Fluorescence microscopy and EM revealed that the cilia of dyf-5 loss-of-function (lf) animals are elongated and are not properly aligned into the amphid channel. Some cilia do enter the amphid channel, but the distal ends of these cilia show accumulation of proteins. Consistent with these observations, we found that six IFT proteins accumulate in the cilia of dyf-5(lf) mutants. In addition, using genetic analyses and live imaging to measure the motility of IFT proteins, we show that dyf-5 is required to restrict kinesin II to the cilia middle segments. Finally, we show that, in dyf-5(lf) mutants, OSM-3 moves at a reduced speed and is not attached to IFT particles. We propose that DYF-5 plays a role in the undocking of kinesin II from IFT particles and in the docking of OSM-3 onto IFT particles.cilia length ͉ dyf-5 ͉ intraflagellar transport C ilia are present on almost every vertebrate cell and have important functions in motility or sensation. Within cilia, structural components and signaling molecules are transported by a specialized system, called intraflagellar transport (IFT) (1-5). Transport from the base of the cilia to the tip (anterograde) is mediated by kinesin-2 motor complexes, whereas dynein motor complexes mediate transport back to the base (retrograde). The nematode Caenorhabditis elegans has 60 ciliated neurons, including eight pairs of amphid neurons exposed to the environment (6). The cilia of these neurons can be divided into a middle segment with nine doublet microtubules and a distal segment with nine singlet microtubules (7). In the middle segments, two distinct kinesin-2 motor complexes mediate anterograde transport, heterotrimeric kinesin II, encoded by klp-11, klp-20, and kap-1 and homodimeric OSM-3 kinesin (8). In the distal segments, transport is mediated by only OSM-3 (8). Live imaging of the movement of these kinesins suggests that kinesin II alone moves at 0.5 m/s, and OSM-3 alone moves at 1.3 m/s, whereas the two motor complexes together move at 0.7 m/s (8). Recently, Pan et al. (9) have shown that these in vivo transport rates can be reconstituted in vitro by using purified kinesin II and OSM-3 motors. Thus far, two proteins have been identified that are required to stabilize IFT complexes transported by both kinesin II and OSM-3, BBS-7 and BBS-8 (10). However, it remains unclear how kinesin II is restricted to the cilia middle segments, whereas OSM-3 is allowed to enter the distal segments and what the functional significance is of this compartmentalization.Recently, Evans e...

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Section: Discussionmentioning

confidence: 86%

mentioning

confidence: 99%

Mutation of the MAP kinase DYF-5 affects docking and undocking of kinesin-2 motors and reduces their speed in the cilia of Caenorhabditis elegans

Burghoorn¹,

Dekkers²,

Rademakers³

et al. 2007

Proc. Natl. Acad. Sci. U.S.A.

116

178

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“…Unfortunately, neither of these kinases was represented in the kinome screen. We chose to further investigate MOK because it had been previously examined and an expression system was established (33,34), while little is known about SgK494. MOK is a serine/threonine kinase of the MAPK superfamily that is expressed in several tissue types.…”

Section: Evaluation Of Cysteine Gatekeeper Targeting Inhibitors For Mokmentioning

confidence: 99%

Chemical genetic strategy for targeting protein kinases based on covalent complementarity

Garske

Peters

Cortesi

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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The conserved nature of the ATP-binding site of the >500 human kinases renders the development of specific inhibitors a challenging task. A widely used chemical genetic strategy to overcome the specificity challenge exploits a large-to-small mutation of the gatekeeper residue (a conserved hydrophobic amino acid) and the use of a bulky inhibitor to achieve specificity via shape complementarity. However, in a number of cases, introduction of a glycine or alanine gatekeeper results in diminished kinase activity and ATP affinity. A new chemical genetic approach based on covalent complementarity between an engineered gatekeeper cysteine and an electrophilic inhibitor was developed to address these challenges. This strategy was evaluated with Src, a proto-oncogenic tyrosine kinase known to lose some enzymatic activity using the shape complementarity chemical genetic strategy. We found that Src with a cysteine gatekeeper recapitulates wild type activity and can be irreversibly inhibited both in vitro and in cells. A cocrystal structure of T338C c-Src with a vinylsulfonamide-derivatized pyrazolopyrimidine inhibitor was solved to elucidate the inhibitor binding mode. A panel of electrophilic inhibitors was analyzed against 307 kinases and MOK (MAPK/MAK/MRK overlapping kinase), one of only two human kinases known to have an endogenous cysteine gatekeeper. This analysis revealed remarkably few offtargets, making these compounds the most selective chemical genetic inhibitors reported to date. Protein engineering studies demonstrated that it is possible to increase inhibitor potency through secondary-site mutations. These results suggest that chemical genetic strategies based on covalent complementarity should be widely applicable to the study of protein kinases. chemical genetics | irreversible inhibitor | SgK494

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“…All MS 2 spectra were collected using the following instrument parameters: one microscan, 3 m/z mass isolation window, default charge state of ϩ10, monoisotopic precursor selection "enabled," and precursor ions in the ϩ1 charge state were excluded from MS 2 analysis. Full automated gain control targets were set to 1e 6 for fourier transform mass spectrometry and 3e 4 for ion trap mass spectrometry. ETD spectra utilized a 30-ms reaction time with a 2-to 4-ms electron transfer reagent injection time with azulene as the electron transfer reagent.…”

Section: Lc-ms/ms Analysis Of Flag-taggedmentioning

confidence: 99%

“…group of the human kinome consists of male germ cell-associated kinase (MAK) 2 (1), ICK/MRK (Intestinal cell kinase/ MAK-related kinase) (2,3) and MAPK/MAK/MRK-overlapping kinase (4). They all share significant homology with MAP kinases in the catalytic domain and contain a MAPK-like TXY motif in the activation T-loop (5,6).…”

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confidence: 99%

Intestinal Cell Kinase (ICK) Promotes Activation of mTOR Complex 1 (mTORC1) through Phosphorylation of Raptor Thr-908

Chapman

Wang

et al. 2012

Journal of Biological Chemistry

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Background:Intestinal cell kinase (ICK), a ubiquitously expressed and highly conserved serine/threonine protein kinase, supports cell proliferation. Results: Phosphorylation of Raptor Thr-908 by ICK is important for full activation of mTORC1. Conclusion: ICK plays an important role in regulating the mTORC1 activity. Significance: These findings provide a novel mechanistic insight into the signaling cascades by which ICK regulates proliferation.

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Molecular cloning and characterization of a novel member of the MAP kinase superfamily

Cited by 78 publications

References 39 publications

Mutation of the MAP kinase DYF-5 affects docking and undocking of kinesin-2 motors and reduces their speed in the cilia of Caenorhabditis elegans

Mutation of the MAP kinase DYF-5 affects docking and undocking of kinesin-2 motors and reduces their speed in the cilia of Caenorhabditis elegans

Chemical genetic strategy for targeting protein kinases based on covalent complementarity

Intestinal Cell Kinase (ICK) Promotes Activation of mTOR Complex 1 (mTORC1) through Phosphorylation of Raptor Thr-908

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