Organization of satellite DNA in the genome of Trypanosoma cruzi

Abstract

Trypanosoma cruzi has about 10(5) Copies of a 195 bp repeat, called satellite DNA, which correspond to about 10% of the total DNA. Since very little is known about satellite distribution and function in this and in many other organisms, we studied the genomic organization of satellite DNA in T. cruzi Y. CL Brener, and Silvio X10 cl1 strains. We found that satellite repeats are present in some, but not all, chromosomes in these strains. We also found that the copy number of the satellite repeats is distinct in each one of the strains. We have estimated that the satellite DNA is four to six times more abundant in Y and CL Brener strains as compared to Silvio X 10 cl1 Despite these differences. the repeated units are organized in a similar way. Based on restriction nuclease digestion analysis, we found that satellite repeats are grouped in clusters comprising about 30 +/- 10 kb. the same cluster size was also found in the different chromosomes, as seen by two-dimensional pulsed field-gel electrophoresis. As evidenced by a fluorescence hybridization assay of spread DNA fibers, we additionally found that these clusters are constrained in specific regions of the chromosomes. Sequence analysis of several repeat units of satellite DNA of the three strains revealed a remarkable similarity with conserved nucleotide blocks separated by more variable regions. Such conserved distribution and common sequence may suggest that these repeated units might have a structural role in the T. cruzi chromosomes. (C) 2003 Elsevier Science B.V. All rights reserved.