Application of high-content analysis to the study of post-translational modifications of the cytoskeleton.
Cytokeratins 8 and 18 have recently been identified as acetylated. The acetylation of other cytoskeletal proteins has been reported as linked to stabilility. As colorectal cells exist bathed in pharmacologically active levels of the HDACi butyrate, we sought to apply state-of-the-art High Content Analytical approaches to identify the effect of butyrate upon the cytoskeleton of colorectal cells. We observed butyrate caused an increase in acetylation at three distinct residues of cytokeratin 8 (K10, K471, and K482) and that the kinetics of altered acetylation were distinct, implying either separate HDACs, or a heirachy of acetylation. This change in acetylation was associated with a breakdown in the cytokeratin cytoskeleton, implying a functional role for cytokeratin acetylation.
Drake
Peter J M
PJ
Human Nutrition Unit, School of Medicine, University of Sheffield, Sheffield S10 2JF, UK.
Griffiths
Gareth J
GJ
Shaw
Lella
L
Benson
Rod P
RP
Corfe
Bernard M
BM
D001331:Automation; D002087:Butyrates / Q000737:chemistry / Q000494:pharmacology*; D018938:Caco-2 Cells; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D015179:Colorectal Neoplasms / Q000188:drug therapy* / Q000378:metabolism*; D003599:Cytoskeleton / Q000187:drug effects* / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D001330:Electronic Data Processing; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007633:Keratins / Q000378:metabolism; D008954:Models, Biological; D011499:Protein Processing, Post-Translational; D040901:Proteomics / Q000379:methods*
D002087:Butyrates; D007633:Keratins