Danesh Moazed's laboratory is working to understand how noncoding RNAs and chromatin-modifying complexes control the transcription of genes and how gene transcription states are inherited during cell division.
Characterization of an RNA Monoadenylation Activity Required for RNAi-Mediated Heterochromatin Assembly
Our lab has recently identified an unusual monoadenylation activity in a noncanonical poly(A) polymerase enzyme that is required for RNAi-mediated heterochromatin formation in Schizosaccharomyces pombe. The activity is allosterically regulated by another protein that is present in the complex. The summer project involves using purified complexes to test the hypothesis that the enzyme stays stably bound to its RNA substrate after addition of the first A and this helps target an associated RNA-dependent RNA polymerase to the RNA substrate to promote double-strand RNA synthesis.
Techniques: protein purification, preparation of T7 polymerase-transcribed RNA, in vitro adenylation assays, gel electrophoresis.