Naslov
Function of an alternative transcript of the Wnt1-inducible signaling pathway protein 1 gene in human renal cells

Autori
Mirjana Polović, Gerald Schwerdt, Michael Gekle

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Skup
95th Annual Meeting of the German Physiological Society (DPG)

Mjesto i datum
Lübeck, Njemačka, 03.-05.3.2016

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
OTA, lncRNA, WISP1

Sažetak
Poster P17-14. The alternative transcript WISP1-OT2 (WNT1 inducible signaling pathway protein 1 overlapping transcript 2) was discovered upon treatment of renal proximal tubule epithelial cells (RPTEC) in culture with the ubiquitous mycotoxin, ochratoxin A (OTA). WISP1-OT2 is 2922 nucleotides long alternative transcript of the WISP1 gene, starting in intron 4 and ending in exon 5. It is detectable predominantly in the nuclear fraction but its function in and interaction with components of renal cells is unknown. Therefore, we studied the interaction of WISP1-OT2 with proteins by miTRAP (microRNA and protein affinity purification) experiments. To get further insights into its function and effects on cells, WISP1-OT2 was overexpressed in human embryonic kidney 293T (HEK293T) cells. Next, we obtained the influence of WISP1-OT2 on protein-coding gene expres- sion (BeadArray microarray technology) and on a whole transcriptome level (RNA sequencing). Northern blot analysis confirmed the expression and length of WISP1-OT2 after OTA exposure. By miTRAP it was shown that WISP1-OT2 can bind to some proteins, mainly splicing factors and IGF2BP proteins as well as to proteins interacting with IGF2BP1 protein. Preliminary results of RNA sequencing indicate that WISP1-OT2 overexpression mostly a) downregulates transcripts encoding adhesion molecules and integral membrane components (e.g. HTR2A and CLDN4) and b) upregulates transcripts coding for immune system components (e.g. CD200 and FCAR) and ion channels (e.g. KCNK3 and TRPC3). The BeadArray microarray technology revealed that the overexpression of WISP1-OT2 affects mainly the expression of genes involved in histone demethylation. In promoter regions of these functionally enriched genes, the most abundant are response elements for SP1 and SP2 transcription factors.mThese results demonstrate that the alternative transcript of the WISP1 gene seems to function as a scaffold for protein complexes. Furthermore, we speculate that it may influence transcription indirectly by pulling away splicing factors from their targets. WISP1-OT2 seems to be involved in regulation of gene expression and splicing but the exact mechanisms are not yet dissected.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA