Nucleic Acids Research, Vol 26, Issue 18 4230-4240, Copyright © 1998 by Oxford University Press
LA Klobutcher, SE Gygax, JD Podoloff, JR Vermeesch, CM Price, CM Tebeau and CL Jahn
During the formation of a new macronucleus in the ciliate Euplotes crassus,
micronuclear chromosomes are reproducibly broken at approximately 10 000
sites. This chromosome fragmentation process is tightly coupled with de
novo telomere synthesis by the telomerase ribonucleoprotein complex,
generating short linear macronuclear DNA molecules. In this study, the
sequences of 58 macronuclear DNA termini and eight regions of the
micronuclear genome containing chromosome fragmentation/telomere addition
sites were determined. Through a statistically based analysis of these
data, along with previously published sequences, we have defined a 10 bp
conserved sequence element (E-Cbs, 5'-HATTGAAaHH-3', H = A, C or T) near
chromosome fragmentation sites. The E-Cbs typically resides within the DNA
destined to form a macronuclear DNA molecule, but can also reside within
flanking micronuclear DNA that is eliminated during macronuclear
development. The location of the E-Cbs in macronuclear-destined versus
flanking micronuclear DNA leads us to propose a model of chromosome
fragmentation that involves a 6 bp staggered cut in the chromosome. The
identification of adjacent macronuclear-destined sequences that overlap by
6 bp provides support for the model. Finally, our data provide evidence
that telomerase is able to differentiate between newly generated ends that
contain partial telomeric repeats and those that do not in vivo.
ARTICLES
Conserved DNA sequences adjacent to chromosome fragmentation and telomere addition sites in Euplotes crassus [published erratum appears in Nucleic Acids Res 1999 Feb 15;27(4):following 1222]
Department of Biochemistry, University of Connecticut Health Center, Farmington, CT 06030, USA. butcher@panda.uchc.edu
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