Periodic Reporting for period 1 - YRNAcleave (Sequence and structural requirements for Y RNA cleavage)
Berichtszeitraum: 2016-08-01 bis 2018-07-31
Y RNAs are ~100 nt long molecules, first discovered owing to their association with the autoimmune proteins Ro60 and La. The four Y RNAs (Y1, Y3, Y4 and Y5) show high evolutionary conversation in the animal kingdom, with humans possessing the full complement of Y RNA genes and some species, such as mouse, only retaining the Y1 and Y3 RNA genes. Y RNAs bind a range of proteins within the cell, notably the toroid shaped protein Ro60 involved in RNA quality control, and the La protein which has a range of implicated functions (Figure 1). Not all Y RNAs however are found within ribonucleoprotein complexes. The non-protein bound Y RNAs have been shown to be essential for the initiation of chromosomal DNA replication by acting as licensing factors in post-midblastula transition (MBT) stage cells.
The overall objective of the project is to understand the process of Y RNA fragmentation in human cells. The project consisted of four specific objectives: Objective 1 Validation of sequencing results for pools 1-3, Objective 2 Mutagenesis of 5' arm of Y5 RNA, Objective 3 Validation of mutations in the Y5 5' sRNA region and Objective 4 Testing ""designer"" mutants. We describe the results and conclusions in the next section.
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We are writing a manuscript at the moment and we are planning to submit it in about 3 months. Trung presented a poster about the work at the Non-coding RNAs conference in Heidelberg in the summer of 2017.