DOI: 10.1101/484246Nov 30, 2018Paper

A novel photoactivatable tool for intermediate filament disruption indicates a role for keratin filaments in early embryogenesis

BioRxiv : the Preprint Server for Biology
Rucha Sanghvi-ShahGregory F. Weber

Abstract

The significance of cytoplasmic intermediate filament proteins has previously been examined largely through various genetic approaches, including knockdown, knockout and transgenic overexpression. Few studies to date have attempted to examine the role of specifically the filamentous intermediate filament network in orchestrating various cell functions. To directly assess the role of the filamentous keratin intermediate filament network in regulation of cellular behavior, we created a PhotoActivatable disruptor of keratin Intermediate Filaments (PA-dIF). This genetically encoded construct consists of a peptide derived from the 2B2 region of Keratin 8 fused to the photosensitive LOV2 domain from Avena sativa phototropin-1. Upon 458 nm photoirradiation, PA-dIF disrupts keratin intermediate filaments in multiple species and cell types. Marked remodeling of the keratin intermediate filament network accompanies collective cellular morphogenetic movements that occur during gastrulation and neurulation in the Xenopus laevis frog embryo. Light-based activation of PA-dIF was able to disrupt keratin intermediate filaments in Xenopus cells and lead to tissue-specific disruption of morphogenetic processes. Altogether our data show a fundame...Continue Reading

Related Concepts

Keratin-13
Embryo
Embryonic Development
Peptides
Xenopus laevis
Intermediate
Gastrulation
Approach
Avena sativa
Neurulation

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