Nucleic Acids Research, 1995, Vol. 23, No. 23 4857-4863
© 1995
Articles |
Deregulated messenger RNA expression during T cell apoptosis
Howard Hughes Medical Institute, New York University Medical Center, Kaplan Cancer Center, Department of Biochemistry 550 First Avenue, New York, NY 10016, USA
*To whom correspondence should be addressed
Received August 28, 1995. Accepted October 20, 1995.
The IL-2 dependent murine cytotoxic T cell line CTLL-2 undergoes programmed cell death when deprived of its specific cytokine. We analyzed the expression of cell cycle related genes after IL-2 deprivation. Here we show that a generalized decrease and re elevation of the levels of mRNA takes place as part of the apoptotic program. The levels of several mRNAs encoding cell cycle functions, including cyclin D2, cyclin D3, cyclin B1, c-myc and max all declined at 1.5–3 h following IL-2 deprivation. Notably, the max mRNA, which was shown to be expressed in proliferating, growth arrested and differentiated cells, is down regulated with the same kinetics as the other mRNAs. Surprisingly, the mRNAs whose levels declined at 1.5–3 h rose again at 10–14 h, a time which closely followed the time of the first detection of apoptotic DNA degradation, at 8 h, but which proceeds actual loss of viability, at 14 h, as measured by trypan blue exclusion. Of all analyzed genes only the expression of the S-phase specific histone H4 gene resists the initial decrease and declines gradually over the course of cell death. Measurement of c-Myc protein synthesis at a late stage of the apoptotic program revealed that the accumulated reinduced mRNA is not translated into protein. Because transcriptional regulation has been shown to be dependent on the chromatin structure, the reinduction may be triggered by relaxation of the chromatin caused by alterations in the chromatin structure of apoptotic cells.
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