Detection of alternative splicing of fibronectin mRNA in a single cell.
Pre-fibronectin mRNA is subject to alternative splicing at three sites, EDA, EDB and IIICS. We analyzed the alternative splicing of fibronectin mRNA in a single cell. Reverse transcription-polymerase chain reaction analyses showed cells that produced a single form of mRNA at each one of these sites as well as cells that produced multiple forms at a given site: for example, some cells produced either the EDA(+) or EDA(-) form of the mRNA and other cells produced both forms. About 80% of the cells produced both (+) and (-) forms of the mRNA at the EDA and EDB sites, and the remaining cells contained either the (+) or (-) form. Five forms of fibronectin mRNA can result from alternative splicing at the IIICS site. Complex combinations of alternative splicing products were observed among the individual cells: there were ten different combinations of mRNA isoforms with respect to the IIICS site. Statistically significant changes in alternative splicing at the IIICS site were observed during cellular senescence.
Kumazaki
T
T
Department of Biochemistry and Biophysics, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan. kumazaki@ue.ipc.hiroshima-u.ac.jp
Mitsui
Y
Y
Hamada
K
K
Sumida
H
H
Nishiyama
M
M
D017398:Alternative Splicing; D002478:Cells, Cultured; D017931:DNA Primers; D005347:Fibroblasts; D005353:Fibronectins / Q000235:genetics*; D006801:Humans; D008957:Models, Genetic; D008967:Molecular Biology / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis*
D017931:DNA Primers; D005353:Fibronectins; D012333:RNA, Messenger