Non-coding RNAs

Many classes of small, mid-sized and long noncoding RNAs, only some of which we will cover MicroRNAs (miRNAs) post-transcriptional gene silencing Piwi-interacting RNAs (piRNAs) control transposable elements and direct DNA methylation at transposable elements Long non-coding RNAs (lncRNAs) appear to direct epigenetic machinery and establish different epigenetic states

Piwi-interacting RNAs (piRNAs)

Best characterised in Drosophila Involved in silencing transposable elements in the germ line and stem cell populations to maintain genomic stability ~24-35 nt in length (longer than miRNAs or siRNAs) Transcribed from uni-directional and bi-directional active transposons or other repeats or transposon/repeat remnants found in piRNA clusters (longer RNAs, then processed) Bound by PIWI proteins (Mili, Miwi1, Miwi2 in mammals, or Piwil1, Piwil2, Piwil4) Different PIWI proteins have non-redundant roles

piRNA biogenesis
Single stranded precursors transcribed from piRNA clusters Primary processing involves shortening Secondary processing - Ping-pong amplification piRNA- PIWI protein complexes allow
1. Degradation of transposon RNA ie. post-transcriptional silencing

piRNA biogenesis
Single stranded precursors transcribed from piRNA clusters Primary processing involves shortening Secondary processing - Ping-pong amplification piRNA- PIWI protein complexes allow
1. Degradation of transposon RNA ie. post-transcriptional silencing 2. DNA methylation of transposon/ repetitive element DNA i.e. transcriptional silencing via RNA directed DNA methylation (MIWI2 imported back to the nucleus)

piRNA and miRNA comparison

miRNAs Dicer dependent, piRNAs Dicer independent miRNA act only in post-transcriptional gene silencing piRNA can act in post-transcriptional gene silencing and piRNAs can act to alter epigenetic state

piRNAs in development and disease

piRNAs are important in the germ line and also in stem cells Mice without various Mili1 and Mili2 proteins are sterile due to impaired spermatogenesis, associated with failure to silence repetitive elements such as LINE1 and IAP elements Mutations in human PIWI proteins associated with infertility PIWI proteins aberrantly expressed in cancer, demonstrating their role outside the germ line, possibly related to stem cells

piRNAs in silencing non-repetitive regions?

piRNAs in pachytene sperm target non-repetitive DNA Active at Rasgrf1 paternally imprinted gene (see later lectures on Imprinting) piRNAs are involved in directing DNA methylation at more loci than just transposable elements Still much to be understood