title: Aberrant heterodimerization of keratin 16 with keratin 6A in HaCaT keratinocytes results in diminished cellular migration.
abstract: Keratin filaments form obligatory heterodimers consisting of one type I and one type II keratin that build the intermediate filaments. In keratinocytes, type II keratin 6 (K6) interacts with type I keratin 16 (K16). We previously showed that the intermediate filament protein K16 is up-regulated in aged human skin. Here, we report that there is an obvious imbalance of K16 to K6 mRNA in in vivo and in vitro aging, which possibly leads to cellular effects. To unveil a possible biological function of K16 overexpression we investigated the migration potential of keratinocytes having up-regulated K16 expression in vitro. Two cell lines were established by transfection of human keratinocytes (HaCaT cells) with K16 or control vectors and subsequent fluorescence-activated cell sorting. By performing migration assays we were able to show a 90% reduction in the migration ability of the K16-overexpressing keratinocytes. In addition, a delay in wound closure associated with K16-overexpressing cells was shown by scratch assays. Transient overexpression of K6A in K16-overexpressing keratinocytes partially corrected the cell-migration defect. By real-time PCR we excluded co-regulation of the annotated interaction partner, K6, in the K16 cell line. Finally, we observed a decreased level of tyrosine phosphorylation in K16-overexpressing cells. Taken together, these data highlight the possibility of a physiological role for K6/K16 heterodimers in keratinocyte cell migration, in addition to the heterodimer's known functions in cell differentiation and mechanical resilience.
authors.affiliation: Division of Molecular Dermatology, Department of Dermatology, Paracelsus Medical University Salzburg, Müllner Hauptstrasse 48, A-5020 Salzburg, Austria. a.trost@salk.at
authors.forename: A
authors.initials: A
authors.lastname: Trost
authors.forename: P
authors.initials: P
authors.lastname: Desch
authors.forename: V
authors.initials: V
authors.lastname: Wally
authors.forename: M
authors.initials: M
authors.lastname: Haim
authors.forename: R H
authors.initials: RH
authors.lastname: Maier
authors.forename: H A
authors.initials: HA
authors.lastname: Reitsamer
authors.forename: H
authors.initials: H
authors.lastname: Hintner
authors.forename: J W
authors.initials: JW
authors.lastname: Bauer
authors.forename: K
authors.initials: K
authors.lastname: Onder
mesh_terms: D002460:Cell Line; D002465:Cell Movement; D016922:Cellular Senescence; D006801:Humans; D053536:Keratin-16 / Q000235:genetics / Q000378:metabolism*; D053553:Keratin-6 / Q000235:genetics / Q000378:metabolism*; D015603:Keratinocytes / Q000378:metabolism / Q000502:physiology*; D010766:Phosphorylation; D055503:Protein Multimerization; D012333:RNA, Messenger / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism
chemical_list: C507569:KRT6A protein, human; D053536:Keratin-16; D053553:Keratin-6; D012333:RNA, Messenger; D014443:Tyrosine