N. Nakamichi et al., Detection and cDNA cloning of H-strand mitochondrial regulatory region RNAs in cultured human cells and human tissues, CYTOTECHNOL, 33(1-3), 2000, pp. 175-188
In the mitochondrion, essential genetic elements for replication and transc
ription are mostly housed within a short segment of its DNA located between
tRNA(Phe) and tRNA(Pro) genes, which is called mitochondrial regulatory re
gion (mrr). RNAs are known to be transcribed from mrr, the structures and t
he functions of which are yet to be fully characterized. We detected ca. 1.
3 kb H-strand transcripts of mrr (mrrH-RNAs), and 0.2 kb L-strand transcrip
ts of mrr (mrrL-RNAs) in various human cultured cells and tissues using dou
ble stranded mrrDNA probes. The steady state levels of mrrL-RNAs were gener
ally high in cultured cells, while they varied among tissues. On the other
hand, the levels of mrrH-RNAs varied among tissues and among cultured cells
. A tendency was observed in these cells and tissues that a high level of m
rrL-RNA is associated with cell proliferation, and a high level of mrrH-RNA
with differentiation. Several cDNA clones to 1.3 kb mrrH-RNA were obtained
from human skeletal muscle polyadenylated RNAs. The 5' terminus of the 1.3
kb RNA was determined to be at nucleotide position 15953 which is immediat
ely downstream of tRNA(T)hr sequence. Polyadenylation site for most of the
clones was demonstrated to be at nucleotide position 576 which is immediate
ly upstream of tRNA(Phe) sequence. The longest cDNA insert obtained was 117
7 bps long spanning from nucleotide positions 15969 to 576 which could code
for a peptide of 76 amino acids. The cDNAs isolated here are the first cDN
A clones reported to human mrrH-RNAs. These results, together with previous
results, further substantiate that polyadenylated mrrH- and mrrL-RNAs are
commonly present at varying levels among human tissues and cells. The 3' en
d sequences of the cloned mrrH-cDNA provides with insights into the mechani
sms of transcription termination. The cDNA clones will provide tools to fur
ther the study of the function of mrr RNAs.