Li Lingyun, Krasnykov Kyrylo, Homolka David, Gos Pascal, Mendel Mateusz, Fish Richard J., Pandey Radha Raman, Pillai Ramesh S. (2022), The XRN1-regulated RNA helicase activity of YTHDC2 ensures mouse fertility independently of m6A recognition, in Molecular Cell
, 82(9), 1678-1690.e12.
Mendel Mateusz, Delaney Kamila, Pandey Radha Raman, Chen Kuan-Ming, Wenda Joanna M., Vågbø Cathrine Broberg, Steiner Florian A., Homolka David, Pillai Ramesh S. (2021), Splice site m6A methylation prevents binding of U2AF35 to inhibit RNA splicing, in Cell
, 184(12), 3125-3142.e25.
Pandey Radha Raman, Delfino Elena, Homolka David, Roithova Adriana, Chen Kuan-Ming, Li Lingyun, Franco Giulia, Vågbø Cathrine Broberg, Taillebourg Emmanuel, Fauvarque Marie-Odile, Pillai Ramesh S. (2020), The Mammalian Cap-Specific m6Am RNA Methyltransferase PCIF1 Regulates Transcript Levels in Mouse Tissues, in Cell Reports
, 32(7), 108038-108038.
Yang Fang, Lan Yemin, Pandey Radha Raman, Homolka David, Berger Shelley L., Pillai Ramesh S., Bartolomei Marisa S., Wang P. Jeremy (2020), TEX15 associates with MILI and silences transposable elements in male germ cells, in Genes & Development
, 34(11-12), 745-750.
Wu Hao, Li Lingyun, Chen Kuan-Ming, Homolka David, Gos Pascal, Fleury-Olela Fabienne, McCarthy Andrew A., Pillai Ramesh S. (2019), Decapping Enzyme NUDT12 Partners with BLMH for Cytoplasmic Surveillance of NAD-Capped RNAs, in Cell Reports
, 29(13), 4422-4434.e13.
Pandey Radha Raman, Homolka David, Olotu Opeyemi, Sachidanandam Ravi, Kotaja Noora, Pillai Ramesh S. (2018), Exonuclease Domain-Containing 1 Enhances MIWI2 piRNA Biogenesis via Its Interaction with TDRD12, in Cell Reports
, 24(13), 3423-3432.e4.
Mendel Mateusz, Chen Kuan-Ming, Homolka David, Gos Pascal, Pandey Radha Raman, McCarthy Andrew A., Pillai Ramesh S. (2018), Methylation of Structured RNA by the m6A Writer METTL16 Is Essential for Mouse Embryonic Development, in Molecular Cell
, 71(6), 986-1000.e11.
Germ cells are entrusted with the task of transmitting the genetic information from one generation to the next. One major threat to germline genome integrity is the mobile genetic elements called transposons. Germline-specific Piwi-interacting RNAs (piRNAs) act as guardians of the genome by silencing transposable elements. The central aims of this proposal are to study how nuclear small RNAs are made and how they function in recruiting DNA methylation to target transposon loci in mammals. Key nucleases involved in piRNA biogenesis are not known. In this proposal, we will characterize the role of two novel nuclease family members in the piRNA biogenesis pathway using biochemical, structural and mouse genetic tools. It will use catalytic-dead mouse mutants of RNA helicases to trap transiently assembled biogenesis and silencing effector machineries in vivo. Since most piRNAs derive from repeat regions, we employ unique artificial piRNA precursors to systematically study piRNA processing steps in insect cell cultures, transgenic fly and mouse models. The use of RNA-crosslinking and deep sequencing strategies will allow us to unambiguously map factor binding sites to these unique reporters. It will aim to biochemically and functionally characterize mammalian piRNA-guided nuclear silencing complexes (RITS). Using artificial reporters and tethering of factors, it will provide direct and unambiguous evidence linking piRNAs to the epigenetic silencing of transposons by DNA methylation in the mammalian genome. Furthermore, it will explore the transgenerational nature of this silencing. Finally, using Cas9 genome editing tools it will examine the role of a set of unique, non-repeat piRNAs whose functions during mouse germ cell development are unknown. Overall, this proposal aims to use interdisciplinary approaches in uncovering the epigenetic role of nuclear germline small RNAs.