Document Type

Article

Major

Molecular & Cell Biology, Applied Mathematical Sciences

Mentor

Prof. Barbara Mellone, Dept. of Molecular & Cell Biology and Institute of Genomics

Disciplines

Genetics and Genomics | Molecular Genetics

Abstract

This study investigates the presence and localization of R-loops at the centromeres of Drosophila melanogaster. R-loops are non-canonical nucleic acid structures that have been implicated in various cellular processes, including genomic instability and accurate chromosome segregation. Although previous research has found R-loops at Drosophila satellite sequences and LTR retrotransposons, their presence and function at the centromeres of a whole organism remain largely unknown. Using IF-FISH (immunofluorescence-fluorescence in situ hybridization), we stained R-loops with the S9.6 antibody. A UAS-rnh1 overexpression line was used to induce the overexpression of RNase H1, an enzyme that resolves R-loops, to confirm the specificity of the S9.6 antibody signal. Our results showed that while inducing rnh1 overexpression did cause a decrease in the frequency of colocalization between S9.6 and CENP-C at all centromeres, this change was only statistically significant for centromere X. Similarly, the decrease in S9.6 and rDNA colocalization was only statistically significant for the X chromosome. The S9.6 signal intensity did not significantly change after induction of rnh1 overexpression. These findings prevent any definitive conclusions regarding the presence of R-loops at Drosophila centromeres.

Comments

The authors thank the Holster Scholars Program, the University of Connecticut Honors Program, and the UConn Foundation for funding.

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