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RNA-seq analysis of budding yeast wild type (BY4741) and prp45(1-169) mutant

ABSTRACT: Prp45 is a budding yeast NineTeen-Complex associated factor, which plays a role in pre-mRNA splicing. In addition to its documented involvement in the second step of splicing, here we show that it is important in the early stages of co-transcriptional spliceosome assembly as well. To determine the overall splicing efficiency of prp45 mutant cells and to reveal how the efficiency depends on the sequences which define introns (whether they are consensual or non-consensual), we performed RNA-seq analysis of prp45(1-169) and corresponding wild-type cells (two biological replicates each). Total RNA was isolated by combining phenol-chlorophorm extraction with MasterPure Yeast RNA Purification Kit (Epicentre). Ribodepletion, library preparation and sequencing were performed by BGI Genomics.

INSTRUMENT(S): Illumina HiSeq 2500

ORGANISM(S): Saccharomyces cerevisiae

SUBMITTER: Petr Folk 

PROVIDER: E-MTAB-5149 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Nineteen complex-related factor Prp45 is required for the early stages of cotranscriptional spliceosome assembly.

Hálová Martina M   Gahura Ondřej O   Převorovský Martin M   Cit Zdeněk Z   Novotný Marian M   Valentová Anna A   Abrhámová Kateřina K   Půta František F   Folk Petr P  

RNA (New York, N.Y.) 20170712 10

Splicing in <i>S. cerevisiae</i> has been shown to proceed cotranscriptionally, but the nature of the coupling remains a subject of debate. Here, we examine the effect of nineteen complex-related splicing factor Prp45 (a homolog of SNW1/SKIP) on cotranscriptional splicing. RNA-sequencing and RT-qPCR showed elevated pre-mRNA levels but only limited reduction of spliced mRNAs in cells expressing C-terminally truncated Prp45, Prp45(1-169). Assays with a series of reporters containing the <i>AMA1</i  ...[more]

PMID: 28701519

Publication: 1/2

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