Gönül Baltacı Nurdan, Koçpınar Enver Fehim, Budak Harun
Department of Molecular Biology and Genetics, Science Faculty, Atatürk University, 25240, Erzurum, Turkey.
Department of Medical Laboratory Techniques, Vocational School of Health Services, Muş Alparslan University, 49250, Muş, Turkey.
Mol Biol Rep. 2021 Nov;48(11):7397-7404. doi: 10.1007/s11033-021-06753-8. Epub 2021 Oct 15.
Carbonic anhydrases (CAs) play a significant role in maintaining pH balance by catalyzing the conversion of carbon dioxide to bicarbonate. The regulation of pH is critical for all living organisms. Although there are many studies in the literature on the biochemical, functional, and structural features of CAs, there is not sufficient information about the epigenetic regulation of CAs.
The lysine acetyltransferase TIP60 (60 kDa Tat-interactive protein) was knocked out specifically in mouse liver using the Cre/loxP system, and knockout rate was shown as 83-88% by Southern blot analysis. The impact of Tip60 on the expression of Ca1, Ca3, and Ca7 was investigated at six Zeitgeber time (ZT) points in the control and liver-specific Tip60 knockout mice (mutant) groups by real-time PCR. In the control group, while Ca1 showed the highest expression at ZT8 and ZT12, the lowest expression profile was observed at ZT0 and ZT20. Hepatic Ca1 displayed robust circadian expression. However, hepatic Ca3 exhibited almost the same level of expression at all ZT points. The highest expression of Ca7 was observed at ZT12, and the lowest expression was determined at ZT4. Furthermore, hepatic Ca7 also showed robust circadian expression. The expression of Ca1 and Ca3 significantly decreased in mutant mice at all time periods, but the expression of Ca7 used as a negative control was not affected.
It was suggested for the first time that Tip60 might be considered a candidate protein in the regulation of the Ca1 and Ca3 genes, possibly by acetylation.
碳酸酐酶（CAs）通过催化二氧化碳转化为碳酸氢盐在维持pH平衡中发挥重要作用。pH调节对所有生物体都至关重要。尽管文献中有许多关于碳酸酐酶的生化、功能和结构特征的研究，但关于碳酸酐酶的表观遗传调控的信息并不充分。
使用Cre/loxP系统在小鼠肝脏中特异性敲除赖氨酸乙酰转移酶TIP60（60 kDa Tat相互作用蛋白），通过Southern印迹分析显示敲除率为83 - 88%。通过实时PCR在对照组和肝脏特异性Tip60敲除小鼠（突变体）组的六个授时时间（ZT）点研究Tip60对Ca1、Ca3和Ca7表达的影响。在对照组中，Ca1在ZT8和ZT12表达最高，而在ZT0和ZT20观察到最低表达谱。肝脏Ca1呈现出强烈的昼夜节律表达。然而，肝脏Ca3在所有ZT点的表达水平几乎相同。Ca7在ZT12表达最高，在ZT4表达最低。此外，肝脏Ca7也呈现出强烈的昼夜节律表达。在所有时间段，突变小鼠中Ca1和Ca3的表达显著降低，但用作阴性对照的Ca7的表达未受影响。
首次提出Tip60可能被视为Ca1和Ca3基因调控中的候选蛋白，可能是通过乙酰化作用。
