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J. Biol. Chem., Vol. 263, Issue 18, 8671-8676, 06, 1988
DS Pasco, KW Boyum, SN Merchant, SC Chalberg and JB Fagan
In both primary cell cultures of rat hepatocytes and in liver, polycyclic
aromatic hydrocarbons (PAHs) were found to influence the accumulation of
the cytochrome P-450c and P-450d mRNAs by both transcriptional and
post-transcriptional mechanisms. Following treatment with PAHs, cytochrome
P-450c mRNA levels increased approximately 100-fold in both hepatocyte
cultures and in liver, while transcription rates, measured by run-on
transcription of isolated nuclei, increased 3-fold in hepatocyte cultures
and 10-fold in liver. The difference in the -fold increases of mRNA level
and transcription rate suggests that post-transcriptional, as well as
transcriptional, mechanisms contributed to the regulation of cytochrome
P-450c mRNA levels. Following treatment with PAHs, cytochrome P-450d mRNA
levels increased 200-fold in hepatocyte cultures and 70-fold in liver,
while transcription rates remained unchanged in hepatocyte cultures and
increased only 1.7-fold in liver. This suggests that post- transcriptional
mechanisms were of primary importance in regulating cytochrome P-450d mRNA
levels. The newly developed hepatocyte primary cell culture system used in
these studies differs from previously reported systems in that the
cytochrome P-450d gene, as well as the cytochrome P-450c gene, were
expressed in response to PAHs. In this cell culture system the regulation
of these two genes was quite similar, although not identical, to that found
in liver. The mechanisms controlling the tissue-specific expression of the
genes encoding cytochromes P-450c and P-450d were also examined. The
cytochrome P-450c mRNA was found in kidney, heart, and lung, as well as in
liver, of PAH- treated rats, while the mature cytochrome P-450d mRNA was
detected only in liver. The substantial increase in cytochrome P-450c mRNA
in kidney in response to beta-napthoflavone was not associated with a
detectable change in the transcription rate of cytochrome P-450c gene,
indicating that cytochrome P-450c mRNA levels must be regulated primarily
post- transcriptionally in kidney. Even though mature cytochrome P-450d
mRNA could not be detected in kidney, the cytochrome P-450d gene was
transcribed at a substantial rate in this tissue; therefore, the lack of
accumulation of mature cytochrome P-450d mRNA in kidney must have been due
to post-transcriptional control.
Transcriptional and post-transcriptional regulation of the genes encoding cytochromes P-450c and P-450d in vivo and in primary hepatocyte cultures
Department of Chemistry, Maharishi International University, Fairfield, Iowa 52556.
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