Nucleic Acids Research, Vol 26, Issue 20 4635-4644, Copyright © 1998 by Oxford University Press
M Yue, KP Reischmann and GM Kapler
The rDNA minichromosomes of Tetrahymena thermophila and Tetrahymena
pyriformis share a high degree of sequence similarity and structural
organization. The T.thermophila 5' non-transcribed spacer (5' NTS) is
sufficient for replication and contains three repeated sequence elements
that are conserved in T.pyriformis , including type I elements, the only
known determinant for replication control. To assess the role of conserved
sequences in replication control, structural and functional studies were
performed on T.pyriformis rDNA. Similar to T.thermophila , replication
initiates exclusively in the 5' NTS, localizing to a 900 bp segment.
Elongating replication forks arrest transiently at one site which bears
strong similarity to a tripartite sequence element present at fork arrest
sites in T.thermophila rDNA. An in vitro type I element binding activity
indistinguishable from the T.thermophila protein, ssA-TIBF, was detected in
T.pyriformis extracts. The respective TIBF proteins bind with comparable
affinity to type I elements from both species, suggesting that in vivo
recognition could cross species boundaries. Despite these similarities, the
T.pyriformis 5' NTS failed to support replication in transformed
T.thermophila cells, suggesting a more complex genetic organization than
previously realized.
ARTICLES
Conserved cis- and trans-acting determinants for replication initiation and regulation of replication fork movement in tetrahymenid species
Department of Medical Biochemistry and Genetics, Texas A&M Health Science Center, College Station, TX 77843-1114, USA.
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