The nonhelical tail domain of keratin 14 promotes filament bundling and enhances the mechanical properties of keratin intermediate filaments in vitro.
Keratin filaments arise from the copolymerization of type I and II sequences, and form a pancytoplasmic network that provides vital mechanical support to epithelial cells. Keratins 5 and 14 are expressed as a pair in basal cells of stratified epithelia, where they occur as bundled arrays of filaments. In vitro, bundles of K5-K14 filaments can be induced in the absence of cross-linkers, and exhibit enhanced resistance to mechanical strain. This property is not exhibited by copolymers of K5 and tailless K14, in which the nonhelical tail domain has been removed, or copolymers of K5 and K19, a type I keratin featuring a short tail domain. The purified K14 tail domain binds keratin filaments in vitro with specificity (kD approximately 2 microM). When transiently expressed in cultured cells, the K14 tail domain associates with endogenous keratin filaments. Utilization of the K14 tail domain as a bait in a yeast two-hybrid screen pulls out type I keratin sequences from a skin cDNA library. These data suggest that the tail domain of K14 contributes to the ability of K5-K14 filaments to self-organize into large bundles showing enhanced mechanical resilience in vitro.
Bousquet
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Department of Biological Chemistry, School of Medicine, Johns Hopkins University, Baltimore, MD 21205, USA.
Ma
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L
Yamada
S
S
Gu
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C
Idei
T
T
Takahashi
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K
Wirtz
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D
Coulombe
P A
PA
D000818:Animals; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000737:chemistry / Q000378:metabolism / Q000648:ultrastructure; D005782:Gels / Q000737:chemistry; D006801:Humans; D007382:Intermediate Filaments / Q000378:metabolism*; D053547:Keratin-14; D053555:Keratin-5; D007633:Keratins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011108:Polymers / Q000737:chemistry; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D005782:Gels; C507540:KRT14 protein, human; C503940:KRT5 protein, human; D053547:Keratin-14; D053555:Keratin-5; D011108:Polymers; D011993:Recombinant Fusion Proteins; D007633:Keratins