Investigating the role of nuclear myosin I in the low serum induced repositioning of chromosome 10 in interphase nuclei

Abstract

The nucleus of mammalian cells has been proven to be highly organised. A recent study on interphase chromosome positioning has identified low serum induced rapid chromosome repositioning. Chromosome 10 initially localised at an intermediate position in normal proliferating human dermal fibroblasts (HDF) was found to relocate to the nuclear periphery 15 minutes after the cells have been incubated in low serum. Whereas chromosome X has remained in a peripheral position. The relocation of chromosome 10 has been shown to be dependant on both actin and myosin functions. In this project we have further investigated the possible role of nuclear myosin I in chromosome 10 repositioning. Using siRNA to block the expression of the nuclear myosin I (NMI) we were able to identify this nuclear myosin as necessary for the rapid repositioning of chromosome 10. Furthermore, using image analysis software we investigated the effect of the NMI knock down on the overall nuclear size and shape. The analysis has revealed that while the nuclear size of normal proliferating cells remained unchanged after the low serum incubation both in cells expressing the NMI and NMI depleted cells, the knock down of the NMI seems to have affected the nuclear shape when the cells were subjected to the serum incubation. On the other hand, the analysis of the chromosome territories area has revealed significant differences in the chromosome territories sizes before and after the low serum incubation, in normal proliferating HDF cells .