Modifications of chromatin structure and gene expression following induced alterations of cellular shape.
In higher eukaryotes cellular shape is a dynamic element which can be altered by external and internal factors (i.e. surface interactions, temperature, ionic strength). Our question was: might modifications of cell shape reflect on nuclear morphology and architecture and hence on chromatin function, in order to represent a mechanism of cell regulation? We altered the shape of cultured fibroblasts by coating the growth substratum with synthetic polymers, which alternatively increased and decreased the adhesiveness. By means of Fluorescence microscopy we analysed the modifications of cell and nucleus architecture induced by the different substrata. Then we used differential scanning calorimetry to investigate if a remodelling of chromatin structure was associated with the induced morphological changes. Finally, we evaluated if the observed modifications of chromatin condensation affect the transcriptional profile. At this stage of the work we focused on just four genes (c-myc, c-fos, c-jun and collagen) and we analysed their expression by dot blot hybridization and RT-PCR. The results confirm that mechanical factors external to the cell, such as the physico-chemical features of the substratum, are able to modulate gene transcription through a remodelling of chromatin structure. Therefore the work supports our starting hypothesis of a regulatory pathway connecting in sequence cellular morphomety/nuclear architecture/chromatin structure/gene expression.
Vergani
Laura
L
Department of Biophysical Sciences and Technologies M&O, School of Medicine, University of Genova, Corso Europa 30, 16132 Genova, Italy. vergani@ibf.unige.it
Grattarola
Myriam
M
Nicolini
Claudio
C
D000818:Animals; D016466:CHO Cells; D002152:Calorimetry, Differential Scanning; D002448:Cell Adhesion / Q000187:drug effects; D002467:Cell Nucleus / Q000502:physiology; D048430:Cell Shape / Q000502:physiology*; D042002:Chromatin Assembly and Disassembly / Q000235:genetics / Q000502:physiology*; D003094:Collagen / Q000235:genetics; D006224:Cricetinae; D005347:Fibroblasts / Q000166:cytology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology*; D008856:Microscopy, Fluorescence; D011108:Polymers / Q000494:pharmacology; D011519:Proto-Oncogenes; D014158:Transcription, Genetic
D011108:Polymers; D003094:Collagen