Marconi M, Sesma A, Rodríguez-Romero JL, González MLR, Wilkinson MD.

Sci Data. 2018 Nov 27;5:180271. doi: 10.1038/sdata.2018.271.

Abstract

Polyadenylation plays an important role in gene regulation, thus affecting a wide variety of biological processes. In the rice blast fungus Magnaporthe oryzae the cleavage factor I protein Rpb35 is required for pre-mRNA polyadenylation and fungal virulence. Here we present the bioinformatic approach and output data related to a global survey of polyadenylation site usage in M. oryzae wild-type and Δrbp35 strains under a variety of nutrient conditions, some of which simulate the conditions experienced by the fungus during part of its infection cycle.

See: https://www.ncbi.nlm.nih.gov/pubmed/30480660

Figure 1: Workflow describing the steps followed to analyze 3′T-fill sequencing data.

After total RNA extraction, Illumina RNA libraries were created following the specifics of the 3′T-Fill protocol⁸. RNA sequencing was then performed on the Illumina HiSeq2000 platform. FastQ reads were processed by removing sequencing adapters and finally aligned onto the reference genome. Alignments were further processed, filtering out low quality mappings, mappings with high A/T content and potential internal priming. Reads were thereafter assigned to overlapping genes up to 400 bp from the annotated gene end. Finally, identified poly(A) sites were clustered together in order to select high-confidence poly(A) sites.