title: The role of Lamin A in cytoskeleton organization in colorectal cancer cells: a proteomic investigation.
abstract: Up-regulated expression of lamin A has been implicated in increased cell invasiveness and mortality in colorectal cancer. Here we use quantitative proteomics to investigate lamin A regulated changes in the cytoskeleton that might underpin increased cell motility. Using siRNA knockdown of lamin A in a model cell line (SW480/lamA) we confirm that the presence of lamin A promotes cell motility. Using an enhanced technique to prepare cytoskeleton fractions in combination with 2D DiGE we were able to accurately and reproducibly detect changes in the representation of protein species within the cytoskeleton as low as 20%. In total 64 protein spots displayed either increased or decreased representation within the cytoskeleton of SW480/lamA cells compared to controls. Of these the identities of 29 spots were determined by mass spectrometry. A majority were multiple forms of three classes of proteins, including components of the actin and IF cytoskeletons, protein chaperones and translation initiation and elongation factors. In particular our data reveal that the representation of tissue transglutaminase 2, which is known to modify elements of the cytoskeleton and is associated with cancer progression, was highly over-represented in the cytoskeleton fraction of SW480/lamA cells. Overall, our data are consistent with changed protein cross-linking and folding that favours the formation of dynamic actin filaments over stress fibres accounting for the altered cell motility properties in SW480/lamA cells.
authors: School of Biological and Biomedical Sciences, Durham University, South Road, Durham DH1 3LE, UK.
authors: Clare R
authors: CR
authors: Foster
authors: Joanne L
authors: JL
authors: Robson
authors: William J
authors: WJ
authors: Simon
authors: Jeremy
authors: J
authors: Twigg
authors: Derek
authors: D
authors: Cruikshank
authors: Robert G
authors: RG
authors: Wilson
authors: Chris J
authors: CJ
authors: Hutchison
mesh_terms: D045744:Cell Line, Tumor; D015179:Colorectal Neoplasms / Q000378:metabolism / Q000473:pathology*; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000502:physiology*; D015180:Electrophoresis, Gel, Two-Dimensional; D019204:GTP-Binding Proteins / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D034904:Lamin Type A / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000502:physiology*; D013058:Mass Spectrometry; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D040901:Proteomics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D011503:Transglutaminases / Q000378:metabolism
chemical_list: D003598:Cytoskeletal Proteins; D034904:Lamin Type A; D034741:RNA, Small Interfering; C000717608:TGM2 protein, human; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins