3′-UTR SNP rs2229611 in G6PC1 affects mRNA stability, expression and Glycogen Storage Disease type-Ia risk

Abstract

The frequency of rs2229611, previously reported in Chinese, Caucasians, Japanese and Hispanics, was investigated for the first time in Indian ethnicity. We analyzed its role in the progression of Glycogen Storage Disease type-Ia (GSD-Ia) and breast cancer. Genotype data on rs2229611 revealed that the risk of GSD-Ia was higher (P=0.0195) with CC compared to TT/TC genotypes, whereas no such correlation was observed with breast cancer cases. We observed a strong linkage disequilibrium (LD) among rs2229611 and other disease causing G6PC1 variants (|D'|=1, r=1). Functional validation performed in HepG2 cells using luciferase constructs showed significant (P<0.05) decrease in expression than wild-type 3'-UTR due to curtailed mRNA stability. Furthermore, AU-rich elements (AREs) mediated regulation of G6PC1 expression characterized using 3'-UTR deletion constructs showed a prominent decrease in mRNA stability. We then examined whether miRNAs are involved in controlling G6PC1 expression using pmirGLO-UTR constructs, with evidence of more distinct inhibition in the reporter function with rs2229611. These data suggests that rs2229611 is a crucial regulatory SNP which in homozygous state leads to a more aggressive disease phenotype in GSD-Ia patients. The implication of this result is significant in predicting disease onset, progression and response to disease modifying treatments in patients with GSD-Ia.

A polymorphism in the transcriptional regulatory region strongly influences ovine FSHR mRNA decay

“…Meanwhile, our results also proved that the c.2327A>G mutation in the 3′‐UTR influences ARE‐mediated mRNA decay of Hu sheep FSHR gene. This is consistent with the polymorphisms in the 3′‐UTR of other ARE‐containing transcripts such as IFNL3 (McFarland et al, ), G6PC1 (Karthi et al, ) and SLC4A4 (Patel et al, ). A more recent report showed that a novel mutation in the 3′‐UTR of human SLC4A4 gene creates an ARE motif and enhances SLC4A4 interaction with the zinc finger protein ZFP36/TTP, an ARE‐mRNA decay‐promoting protein, thereby leading to destabilization and rapid degradation of the SLC4A4 transcript (Patel et al, ).…”

“…Furthermore, both ARE1 and ARE2 motifs in the 3′‐UTR are all involved in mRNA decay of Hu sheep FSHR gene (Figure g). Together, our findings further demonstrated the importance of AREs in the regulation of expression of the ARE‐containing transcripts (Chen & Shyu, ; Karthi et al, ; Patel et al, ). Meanwhile, our results also proved that the c.2327A>G mutation in the 3′‐UTR influences ARE‐mediated mRNA decay of Hu sheep FSHR gene.…”

“…Meanwhile, our results also proved that the c.2327A>G mutation in the 3′-UTR influences ARE-mediated mRNA decay of Hu sheep FSHR gene. This is consistent with the polymorphisms in the 3′-UTR of other ARE-containing transcripts such as IFNL3 (McFarland et al, 2014), G6PC1 (Karthi et al, 2017) and SLC4A4 (Patel et al, 2017).…”

“…In contrast to the 5′ regulatory region, polymorphism in the 3′‐UTR is rapidly becoming a new hot spot in regulating gene transcription. Recently, it has been reported that several mutations in the 3′‐UTR could regulate gene transcription, such as rs4803217, rs3088442 and rs2229611 in IFNL3 (McFarland et al, ), SLC22A3 (Li et al, ) and G6PC1 (Karthi et al, ), respectively. However, such mutations are mainly reported in cancer‐related genes or human disease‐related genes, and less in livestock.…”

“…AREs are important cis-acting elements that were initially found in the 3′-UTR of the mRNA, which promote mRNA decay by interacting with ARE-binding proteins, such as HuR and TTP (Bakheet, Hitti, & Khabar, 2018). Interestingly, we had previously found that transcripts (Chen & Shyu, 1995;Karthi et al, 2017;Patel et al, 2017).…”