Profiling RNA‐Associated Proteins for Vasopressin‐Mediated Translation in Kidney Collecting Duct Cells

Abstract

Vasopressin‐mediated gene expression changes in the renal collecting duct has been intensively studied over the last decade. Prior studies were focused on transcriptional regulation as a mechanism of regulation of transport by vasopressin. However, post‐transcriptional regulation at the RNA level has not been studied. To identify vasopressin‐mediated RNA modulators, 1) a comprehensive list of RNA‐associated proteins was curated, and 2) vasopressin‐regulated genes from this list were identified using RNA‐seq in mouse mpkCCD cells exposed to vasopressin for various times.The curated database of RNA‐associated proteins was generated by mining information from multiple databases as follows. RNA processing‐related proteins were identified from Gene Ontology (GO) ‘Molecular Function’ and ‘Biological Process’, KEGG pathway, RNA modification database and Reactome. In addition, proteins that have RNA‐binding domains were identified from five different databases (RBPDB, ATtRACT, RBPmap, InterPro and UniProt). A total of 1369 proteins are listed in the current database of RNA‐associated proteins (https://hpcwebapps.cit.nih.gov/ESBL/Database/RNAprocessing_Proteins/). Among these proteins, 646 proteins have known RNA‐binding domain structures and 723 proteins were identified with annotated molecular functions related to RNA processing. 371 proteins were identified by both criteria.RNA‐seq was carried out in mpkCCD cells treated with the vasopressin analog dDAVP (10−10 M) for 3, 6 and 12 h. Relative transcript abundance was estimated using transcripts per million (TPM) and change of transcript abundance was presented as Log2 of the dDAVP:vehicle TPM ratio at each time point. Interestingly, we found that transcript abundance of genes coding for ribosomal proteins (60S ribosomal proteins and 40S ribosomal proteins) and translation initiation proteins (eukaryotic translation initiation factors) were extensively changed by dDAVP at all time points. These results indicate that vasopressin‐mediated gene expression includes changes mediated by abundance changes of translation‐associated proteins. Beyond this, the current study provides new resources for further study of RNA‐associated processes in the kidney.Support or Funding InformationThe work was supported by the Division of Intramural Research, National Heart, Lung, and Blood Institute projects ZIA‐HL001285 (to MAK) and ZIA‐HL006129 (to MAK).This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.