Long noncoding RNA YLB regulates expression of multiple genes in subtelomeric regions
Objective To discover a novel long noncoding RNA YLL066W-B (referred to as YLB), whose expression can be regulated by a ubiquitin ligases E3, Huwel/Tom1, and further investigate the regulatory effects of YLB on expression of multiple subtelomeric genes. Methods Yeast strains (including Tom1△, YLB-HA, HA-YLB, pYES2-HA-YLB and YLB△) were constructed according to the principle of PCR-based tagging of yeast genes. The effects of Tom1 deletion on gene expression were analyzed by real-time PCR and DNA microarray. The protein levels were detected by Western blot. We further performed quantitative real time-PCR to analyze the inlfuence of YLB on expression of multiple subtelomerical genes.Results We found that deletion of Tom1 in yeast could affect the expression of multiple genes and greatly up-regulated the expression of YLB, which is implicated in cell cycle regulation. By analyzing its nucleotide sequence(171 bp)and detecting protein expression, we speculate that the transcriptional product of YLB is a long noncoding RNA (lncRNA). Although YLB is not homologous to any protein-encoding sequences by NCBI blast, it is homologous to the upstream or downstream regions of the open reading frame of several subtelomerically-encoded genes, including those from pau family and DNA helicase Yrf family. Thus, it is possible that YLB is involved in the regulation of these subtelomerically-encoded genes. Accordingly, deletion of YLB markedly up-regulated the mRNA levels of Yrf1-4, pau4 and pau22, whereas over-expression of YLB greatly down-regulated their expression.Conclusion We have discovered the novel lncRNA YLB. The expression of YLB could be negatively regulated by Tom1, and YLB could regulate the expression of multiple subtelomeric genes.
NETL
NSTL
万方数据
维普
