Clemson University
--J. Wang, J. Barth and A. Abbott
The study of mitochondrial gene expression in higher plants is hampered
by the inability to easily manipulate the system; therefore, we have chosen
to study maize lines which have naturally occurring variation in gene expression.
Comparative molecular analyses of these expression variants with normal
systems will define components essential to the process of mitochondrial
gene transcription, processing and translation. Using Northern hybridization
analysis, mitochondrial RNA's from diverse cytoplasmic backgrounds have
been screened for variations in expression of a number of cloned genes
or unassigned reading frames. Most sources of N cytoplasm have identical
transcript patterns for the probes examined (N. H. Walker et al., Theor.
Appl. Genet. 74:531, 1987). One line, however, B37N, shows an altered transcript
pattern for the URF-25 reading frame originally described by R. E. Dewey
et al. (Cell 44:439, 1986). In this line the mature URF-25 transcript is
not present at the 2200 nucleotide size seen in B73N cytoplasm and all
other lines examined. The major transcript for this URF in B37N is found
at the probable precursor size of approximately 3800 nucleotides. Using
flanking and reading frame specific probes in Northern hybridization analysis
the putative precursor transcript has been characterized in B37N and B73N
mitochondrial RNA's. These transcripts appear to be identical. One plausible
explanation for the lack of a 2200 base transcript in B37N is the loss
of a processing sequence on either the 5' or 3' end of the transcript.
To examine this possibility clones containing URF-25 and flanking sequences
from B37N and B73N mitochondrial DNA have been isolated and comparative
restriction fragment analysis has been carried out. There does not appear
to be any gross difference in the structural organization of this transcription
unit in either cytoplasm. Using cloned probes, the 5' end of the mature
transcript has been mapped and is being sequenced in both B37N and B73N
to search for minor sequence changes. Similar experiments will be done
on the 3' end. We are also testing the possibility that nuclear background
may influence the expression of this URF. These experiments will delimit
salient features involved in mature transcript size determination in higher
plant mitochondria.
Return to the MNL 63 On-Line Index
Return to the Maize Newsletter Index
Return to the MaizeGDB Homepage