Citron kinase

Citron Rho-interacting kinase is an enzyme that in humans is encoded by the CIT gene.

CIT
Identifiers
Aliases	CIT, CRIK, STK21, citron rho-interacting serine/threonine kinase, MCPH17, CITK
External IDs	OMIM: 605629; MGI: 105313; HomoloGene: 21404; GeneCards: CIT; OMA:CIT - orthologs
Gene location (Human) Chr. Chromosome 12 (human) Band 12q24.23 Start 119,685,791 bp End 119,877,320 bp
Gene location (Mouse) Chr. Chromosome 5 (mouse) Band 5|5 F Start 115,983,337 bp End 116,147,006 bp
RNA expression pattern Bgee Human Mouse (ortholog) Top expressed in lateral nuclear group of thalamus right frontal lobe sural nerve primary visual cortex Brodmann area 9 caudate nucleus putamen postcentral gyrus cingulate gyrus anterior cingulate cortex Top expressed in medial dorsal nucleus lateral geniculate nucleus medial geniculate nucleus basilar part of occipital bone zygote substantia nigra secondary oocyte lumbar subsegment of spinal cord medial vestibular nucleus rib More reference expression data BioGPS More reference expression data
Gene ontology Molecular function transferase activity nucleotide binding protein kinase activity PDZ domain binding SH3 domain binding scaffold protein binding metal ion binding kinase activity protein serine/threonine kinase activity protein binding ATP binding Cellular component cytoplasm cytosol membrane soma plasma membrane Golgi cisterna cleavage furrow midbody nucleus cytoskeleton actin cytoskeleton intracellular anatomical structure Biological process cell differentiation intracellular signal transduction phosphorylation nervous system development generation of neurons positive regulation of cytokinesis multicellular organism development cell division protein phosphorylation cell cycle G2/M transition of mitotic cell cycle regulation of actin polymerization or depolymerization Golgi organization mitotic cytokinesis neuron apoptotic process mitotic cell cycle liver development peptidyl-threonine phosphorylation cortical actin cytoskeleton organization actomyosin structure organization Sources:Amigo / QuickGO
Orthologs Species Human Mouse Entrez 11113 12704 Ensembl ENSG00000122966 ENSMUSG00000029516 UniProt O14578 P49025 RefSeq (mRNA) NM_001206999 NM_007174 NM_007708 RefSeq (protein) NP_001193928 NP_009105 NP_031734 Location (UCSC) Chr 12: 119.69 – 119.88 Mb Chr 5: 115.98 – 116.15 Mb PubMed search
Wikidata
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Structure

Citron is a 183 kDa protein that contains a C6H2 zinc finger, a PH domain, and a long coiled-coil forming region including 4 leucine zippers and a rho / rac binding site. It was discovered as a rho/rac effector in 1995, interacting only with the GTP bound forms of rho and rac 1. Displaying a distinctive protein organization, this protein defines a separate class of rho partners. Using a cloning approach based on the polymerase chain reaction (PCR), a splice variant of citron, citron kinase (citron-K) has been identified with an alternative amino terminus. This N-terminal extension contains a protein kinase domain that has approximately 50% sequence identity to the sequences of ROCK, ROK, myotonic dystrophy protein kinase (MDPK) and the CDC42 effector known as MRCK or GEK. Citron kinase, which resembles the ROCK family of kinases and by comparison to it, is therefore a multiple domain protein containing an N-terminal kinase domain, an internal coiled-coil (CC) domain with Rho/Rac interacting site, and a C-terminal region consisting of a Zn finger, a pleckstrin homology (PH) domain, a Citron homology domain (CNH), a putative SH3 binding domain, and a PDZ-targeting motif. Its fly (Drosophila) ortholog is called Sticky. the importance of different domains of citron-K in its localization at different stages is discussed below.

Tissue distribution, localization and dynamics

Investigating the tissue distribution of the citron isoforms with and without the kinase domain, it has been shown that the non-kinase form is restricted to the brain region while the kinase form is widely expressed. Immunofluorescence analyses determined the localization of citron-K and its behavior during cytokinesis. Citron-K first appeared at the equatorial cortex in anaphase, concentrated at the cleavage furrow in early telophase, accumulated in the middle of the intercellular bridge with full ingression of the cleavage furrow in mid telophase, and formed a ring-like structure in the midbody in late telophase, with negligible turnover rate at the midbody. In other words, the protein is much less dynamic at the midbody. Using a series of deletions, it was observed that distinct regions of Citron-K CC (coiled- coil) domain differentially regulate the localizations of Citron-K during cytokinesis. The C terminal part of the CC domain localized at the cleavage furrow and the midbody while the N-terminal part of the CC domain localized at the central spindle in early telophase and on outer region of the midbody in late telophase.

Function

As previously mentioned, citron-K was believed to act in cytokinesis. Its depletion impairs maintenance of the midbody and its overexpression in HeLa cells rendered host cells multinucleated. Cytokinesis failure of Citron-K-depleted cells occurred after full ingression of the cleavage furrow, at the abscission stage. Microtubule disassembly was not seen in any of Citron-K depleted cells with cytokinesis failure. The dominant mode of failure was the inability of daughter cells, which are connected with a shorter intercellular bridge, to separate well. As the midbody microtubules were displaced from the center toward either of the two daughter cells, the two cells fused again with the microtubules absorbed into that daughter cell. To sum the process, Citron-K is important to keep proper structure of the midbody which holds the intercellular bridge microtubules between the two daughter cells and is thus required for successful transition from constriction to abscission. In molecular terms, citron-K depletion impaired the accumulation of 3 key proteins: Rho, Anillin and septins (specifically septin 6 and 7) in the intercellular bridge in mid–late telophase, which in previous stages early to mid-telophase was found to co-localize with them.

Interactions

CIT (gene) has been shown to interact with RHOB and RHOA.

Citron-K or its fly orthologue Sticky has been suggested to interact with several molecules in cytokinesis such as Kinesin-3 (KIF14), actin, myosin light chain, and anillin.

KIF14

The N- terminal of the coiled- coil domain of Citron-K directly interacts with the 2nd coiled-coil domain of KIF14. The localization of KIF14 and citron kinase to the central spindle and midbody is codependent, and they form a complex depending on the activation state of citron kinase. This suggests that the regulation of the interaction between KIF14 and Citron-K is important for Citron-K localization to exert its function but this interaction alone cannot accomplish cytokinesis fully.

ASPM

ASPM (abnormal spindle-like microcephaly associated) is localized to the spindle pole, and is essential for maintaining proliferative cell division. It has been reported that ASPM also localizes to the midbody ring in mammalian cells. This was due to the observed differential localization of the N-terminal and C-terminal regions of ASPM within mitotic cells to either spindle poles or to midbodies, respectively. Since, ASPM co-localizes with Citron-K at the midbody ring in HeLa cells and in developing neocortex, it has been proposed that ASPM may function to coordinate spindle rotation with localization of abscission through interaction with Citron-K.

Actomyosin filaments

In many organisms, the force that drives furrow ingression is the assembly and contraction of actomyosin filaments that often form a contractile ring. The contractile ring is a very dynamic structure in which actomyosin filaments are continuously assembled and disassembled. The small GTPase RhoA has been shown to be implicated, controlling CR assembly and dynamics during cytokinesis. This GTPase cycles between an inactive GDP-bound form and an active GTP-bound form, and this RhoA flux seems important for contractile ring dynamics. In drosophila, Sti (Sticky, ortholog of Citron-K) localizes to the cleavage furrow via association of a predicted coiled-coil region with actin and myosin. However, Sti depletion perturbs RhoA localization and causes excessive accumulation of phosphorylated MRLC (myosin regulatory light chain) at the cleavage site in late cytokinesis. Sti is believed to maintain correct RhoA localization at the cleavage site, which is in turn important for proper contractile ring organization at the end of cytokinesis.

Anillin

The scaffold protein anillin is one of the most crucial partners of RhoA during cytokinesis and plays a fundamental role in the assembly and stabilization of the contractile ring by interacting with RhoA, septins, F-actin, myosin II, and mDia2 and it has been shown that its depletion results in cleavage furrow instability. Citron-K is capable of physically and functionally interacting with the actin-binding protein anillin. Like active RhoA, anillin is also displaced from the midbody in Citron-K-depleted cells. The overexpression of Citron-K and of anillin leads to abscission delay. These results stress that Citron-K is a crucial abscission regulator that may promote midbody stability through active RhoA and anillin.

Clinical implications

Citron-K is expressed during neurogenesis and plays important roles in neuronal progenitor cell division. Recessive mutations in Citron-K cause severe microcephaly both in rats and mice. In humans and rodents, loss of Citron-K expression results in defects in neurogenic cytokinesis. Similarly in Drosophila, RNAi knockdown of Citron-K results in a failure of cellular abscission. CIT is associated with microlissencephaly.

External links

• Human CIT genome location and CIT gene details page in the UCSC Genome Browser.

Further reading

• Madaule P, Furuyashiki T, Reid T, Ishizaki T, Watanabe G, Morii N, Narumiya S (Dec 1995). "A novel partner for the GTP-bound forms of rho and rac". FEBS Letters. 377 (2): 243–8. doi:10.1016/0014-5793(95)01351-2. PMID 8543060. S2CID 39746553.
• Zhang W, Vazquez L, Apperson M, Kennedy MB (Jan 1999). "Citron binds to PSD-95 at glutamatergic synapses on inhibitory neurons in the hippocampus" (PDF). The Journal of Neuroscience. 19 (1): 96–108. doi:10.1523/JNEUROSCI.19-01-00096.1999. PMC 6782379. PMID 9870942.
• Nagase T, Ishikawa K, Suyama M, Kikuno R, Hirosawa M, Miyajima N, Tanaka A, Kotani H, Nomura N, Ohara O (Feb 1999). "Prediction of the coding sequences of unidentified human genes. XIII. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro". DNA Research. 6 (1): 63–70. doi:10.1093/dnares/6.1.63. PMID 10231032.
• Husi H, Ward MA, Choudhary JS, Blackstock WP, Grant SG (Jul 2000). "Proteomic analysis of NMDA receptor-adhesion protein signaling complexes". Nature Neuroscience. 3 (7): 661–9. doi:10.1038/76615. hdl:1842/742. PMID 10862698. S2CID 14392630.
• Di Cunto F, Imarisio S, Hirsch E, Broccoli V, Bulfone A, Migheli A, Atzori C, Turco E, Triolo R, Dotto GP, Silengo L, Altruda F (Oct 2000). "Defective neurogenesis in citron kinase knockout mice by altered cytokinesis and massive apoptosis". Neuron. 28 (1): 115–27. doi:10.1016/S0896-6273(00)00090-8. PMID 11086988. S2CID 15653045.
• Liu H, Di Cunto F, Imarisio S, Reid LM (Jan 2003). "Citron kinase is a cell cycle-dependent, nuclear protein required for G2/M transition of hepatocytes". The Journal of Biological Chemistry. 278 (4): 2541–8. doi:10.1074/jbc.M210391200. PMID 12411428.
• Ozeki Y, Tomoda T, Kleiderlein J, Kamiya A, Bord L, Fujii K, Okawa M, Yamada N, Hatten ME, Snyder SH, Ross CA, Sawa A (Jan 2003). "Disrupted-in-Schizophrenia-1 (DISC-1): mutant truncation prevents binding to NudE-like (NUDEL) and inhibits neurite outgrowth". Proceedings of the National Academy of Sciences of the United States of America. 100 (1): 289–94. doi:10.1073/pnas.0136913100. PMC 140954. PMID 12506198.
• Irobi J, Van Impe K, Seeman P, Jordanova A, Dierick I, Verpoorten N, Michalik A, De Vriendt E, Jacobs A, Van Gerwen V, Vennekens K, Mazanec R, Tournev I, Hilton-Jones D, Talbot K, Kremensky I, Van Den Bosch L, Robberecht W, Van Vandekerckhove J, Van Broeckhoven C, Gettemans J, De Jonghe P, Timmerman V (Jun 2004). "Hot-spot residue in small heat-shock protein 22 causes distal motor neuropathy". Nature Genetics. 36 (6): 597–601. doi:10.1038/ng1328. PMID 15122253.
• Lyons-Warren A, Chang JJ, Balkissoon R, Kamiya A, Garant M, Nurnberger J, Scheftner W, Reich T, McMahon F, Kelsoe J, Gershon E, Coryell W, Byerley W, Berrettini W, Depaulo R, McInnis M, Sawa A (Sep 2005). "Evidence of association between bipolar disorder and Citron on chromosome 12q24". Molecular Psychiatry. 10 (9): 807–9. doi:10.1038/sj.mp.4001703. PMID 15983625.
• Gruneberg U, Neef R, Li X, Chan EH, Chalamalasetty RB, Nigg EA, Barr FA (Jan 2006). "KIF14 and citron kinase act together to promote efficient cytokinesis". The Journal of Cell Biology. 172 (3): 363–72. doi:10.1083/jcb.200511061. PMC 2063646. PMID 16431929.
• Nousiainen M, Silljé HH, Sauer G, Nigg EA, Körner R (Apr 2006). "Phosphoproteome analysis of the human mitotic spindle". Proceedings of the National Academy of Sciences of the United States of America. 103 (14): 5391–6. Bibcode:2006PNAS..103.5391N. doi:10.1073/pnas.0507066103. PMC 1459365. PMID 16565220.
• Olsen JV, Blagoev B, Gnad F, Macek B, Kumar C, Mortensen P, Mann M (Nov 2006). "Global, in vivo, and site-specific phosphorylation dynamics in signaling networks". Cell. 127 (3): 635–48. doi:10.1016/j.cell.2006.09.026. PMID 17081983. S2CID 7827573.