{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,21]],"date-time":"2026-01-21T08:29:53Z","timestamp":1768984193631,"version":"3.49.0"},"reference-count":33,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"6","license":[{"start":{"date-parts":[[2015,8,25]],"date-time":"2015-08-25T00:00:00Z","timestamp":1440460800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2015,8,25]],"date-time":"2015-08-25T00:00:00Z","timestamp":1440460800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01 DK47918"],"award-info":[{"award-number":["R01 DK47918"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["K01 DK093776"],"award-info":[{"award-number":["K01 DK093776"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["DK34933"],"award-info":[{"award-number":["DK34933"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["lww.com","ovid.com"],"crossmark-restriction":true},"short-container-title":["Hepatology"],"published-print":{"date-parts":[[2015,12]]},"abstract":"<jats:p>Keratins, among other cytoskeletal intermediate filament proteins, are mutated at a highly conserved arginine with consequent severe disease phenotypes due to disruption of keratin filament organization. We screened a kinase inhibitor library, using A549 cells that are transduced with a lentivirus keratin 18 (K18) construct, to identify compounds that normalize filament disruption due to K18 Arg90Cys mutation at the conserved arginine. High\u2010throughput screening showed that PKC412, a multikinase inhibitor, ameliorated K18 Arg90Cys\u2010mediated keratin filament disruption in cells and in the livers of previously described transgenic mice that overexpress K18 Arg90Cys. Furthermore, PKC412 protected cultured A549 cells that express mutant or wild\u2010type K18 and mouse livers of the K18 Arg90Cys\u2010overexpressing transgenic mice from Fas\u2010induced apoptosis. Proteomic analysis of proteins that associated with keratins after exposure of K18\u2010expressing A549 cells to PKC412 showed that nonmuscle myosin heavy chain\u2010IIA (NMHC\u2010IIA) partitions with the keratin fraction. The nonmuscle myosin\u2010IIA (NM\u2010IIA) association with keratins was confirmed by immune staining and by coimmunoprecipitation. The keratin\u2013myosin association is myosin dephosphorylation\u2013dependent; occurs with K8, the obligate K18 partner; is enhanced by PKC412 in cells and mouse liver; and is blocked by hyperphosphorylation conditions in cultured cells and mouse liver. Furthermore, NMHC\u2010IIA knockdown inhibits PKC412\u2010mediated normalization of K18 R90C filaments. <jats:italic toggle=\"yes\">Conclusion<\/jats:italic>: The inhibitor PKC412 normalizes K18 Arg90Cys mutation\u2010induced filament disruption and disorganization by enhancing keratin association with NM\u2010IIA in a myosin dephosphorylation\u2013regulated manner. Targeting of intermediate filament disorganization by compounds that alter keratin interaction with their associated proteins offers a potential novel therapeutic approach for keratin and possibly other intermediate filament protein\u2013associated diseases.(H<jats:sc>epatology<\/jats:sc> 2015;62:1858\u20131869)<\/jats:p>","DOI":"10.1002\/hep.27965","type":"journal-article","created":{"date-parts":[[2015,6,30]],"date-time":"2015-06-30T23:16:46Z","timestamp":1435706206000},"page":"1858-1869","update-policy":"https:\/\/doi.org\/10.1097\/lww.0000000000001000","source":"Crossref","is-referenced-by-count":31,"title":["PKC412 normalizes mutation\u2010related keratin filament disruption and hepatic injury in mice by promoting keratin\u2013myosin binding"],"prefix":"10.1097","volume":"62","author":[{"given":"Raymond","family":"Kwan","sequence":"first","affiliation":[{"name":"Departments of Molecular & Integrative Physiology and of Medicine,University of Michigan,Ann Arbor,MI"},{"name":"VA Ann Arbor Healthcare System,Ann Arbor,MI"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lu","family":"Chen","sequence":"additional","affiliation":[{"name":"Departments of Molecular & Integrative Physiology and of Medicine,University of Michigan,Ann Arbor,MI"},{"name":"Infectious Diseases Department,Ruijin Hospital, Shanghai Jiao Tong University Medical School,Shanghai,People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Koksun","family":"Looi","sequence":"additional","affiliation":[{"name":"Departments of Molecular & Integrative Physiology and of Medicine,University of Michigan,Ann Arbor,MI"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Guo\u2010Zhong","family":"Tao","sequence":"additional","affiliation":[{"name":"Department of Surgery,Stanford University,Palo Alto,CA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sujith V.","family":"Weerasinghe","sequence":"additional","affiliation":[{"name":"Departments of Molecular & Integrative Physiology and of Medicine,University of Michigan,Ann Arbor,MI"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Natasha T.","family":"Snider","sequence":"additional","affiliation":[{"name":"Departments of Molecular & Integrative Physiology and of Medicine,University of Michigan,Ann Arbor,MI"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mary Anne","family":"Conti","sequence":"additional","affiliation":[{"name":"The Laboratory of Molecular Cardiology,National Heart, Lung, and Blood Institute, National Institutes of Health,Bethesda,MD"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Robert S.","family":"Adelstein","sequence":"additional","affiliation":[{"name":"The Laboratory of Molecular Cardiology,National Heart, Lung, and Blood Institute, National Institutes of Health,Bethesda,MD"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qing","family":"Xie","sequence":"additional","affiliation":[{"name":"Infectious Diseases Department,Ruijin Hospital, Shanghai Jiao Tong University Medical School,Shanghai,People's Republic of China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M. Bishr","family":"Omary","sequence":"additional","affiliation":[{"name":"Departments of Molecular & Integrative Physiology and of Medicine,University of Michigan,Ann Arbor,MI"},{"name":"VA Ann Arbor Healthcare System,Ann Arbor,MI"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"276","published-online":{"date-parts":[[2015,8,25]]},"reference":[{"key":"hep27965-bib-0001-20241017","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1016\/j.tig.2012.06.001","article-title":"Nuclear lamin functions and disease","volume":"28","year":"2012","journal-title":"Trends Genet"},{"key":"hep27965-bib-0002-20241017","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1146\/annurev.bi.63.070194.002021","article-title":"Intermediate filaments: structure, dynamics, function, and disease","volume":"63","year":"1994","journal-title":"Annu Rev 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