SOX9 and SRY binding sites on mouse mXYSRa/Enh13 enhancer redundantly regulate <i>Sox9</i> expression to varying degrees
SOX9 and SRY binding sites on mouse mXYSRa/Enh13 enhancer redundantly regulate <i>Sox9</i> expression to varying degrees
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小鼠 mXYSRa/Enh13 增强子上的 SOX9 和 SRY 结合位点不同程度地冗余调节 <i>Sox9</i> 表达
DOI：
10.1093/hmg/ddac184
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发表时间：
2022
影响因子：
3.5
通讯作者：
Takada Shuji
中科院分区：
文献类型：
--
作者：
Ogawa Yuya;Terao Miho;Tsuji-Hosokawa Atsumi;Tsuchiya Iku;Hasegawa Midori;Takada Shuji
Sox 9在哺乳动物睾丸形成中起重要作用。据报道，睾丸中的基因表达受增强子调控。其中，mXYSRa/Enh 13-位于转录起始位点的远上游-起着关键作用，其中其缺失导致小鼠中完全的雄性到雌性的性别逆转。已经提出SOX 9和SRY的结合位点（BS）与mXYSRa/Enh 13相关，SRY是Y染色体上的性别决定基因。它们作为增强子起作用，因此这些序列在进化上是保守的，并且先前已经证明了SOX 9和SRY与mXYSRa/Enh 13的体内结合。然而，他们的精确vivofunctions还没有被检查的日期。为此，本研究产生了在SOX 9和SRY BS上具有替换的小鼠，以揭示它们的体内功能。SOX 9和SRY BS的纯合突变体与XY雄性小鼠无法区分，而双突变体具有小睾丸，这表明这些功能是多余的，并且mXYSRa/Enh 13上存在另一个功能序列，因为mXYSRa/Enh 13缺失小鼠是XY雌性小鼠。此外，大多数在S 0X 9 BS和SRY BS中具有取代的半合子小鼠分别是雌性和雄性，这表明S 0X 9 BS对SRY BS的贡献更多，以使mXYSRa/Enh 13发挥功能。利用体外试验验证了SOX 9和SRY通过这些BS的累加效应。结论：SOX 9 BS和SRY BS在体内的功能是冗余的，mXYSRa/Enh 13中至少应存在一个以上的功能序列。
Sox9plays an essential role in mammalian testis formation. It has been reported that gene expression in the testes is regulated by enhancers. Among them, mXYSRa/Enh13—which is located at far upstream of the transcription start site—plays a critical role, wherein its deletion causes complete male-to-female sex reversal in mice. It has been proposed that the binding sites (BSs) of SOX9 and SRY, the latter of which is the sex determining gene on the Y chromosome, are associated with mXYSRa/Enh13. They function as an enhancer, whereby the sequences are evolutionarily conserved andin vivobinding of SOX9 and SRY to mXYSRa/Enh13 has been demonstrated previously. However, their precisein vivofunctions have not been examined to date. To this end, this study generated mice with substitutions on the SOX9 and SRY BSs to reveal theirin vivofunctions. Homozygous mutants of SOX9 and SRY BS were indistinguishable from XY males, whereas double mutants had small testes, suggesting that these functions are redundant and that there is another functional sequence on mXYSRa/Enh13, since mXYSRa/Enh13 deletion mice are XY females. In addition, the majority of hemizygous mice with substitutions in SOX9 BS and SRY BS were female and male, respectively, suggesting that SOX9 BS contributes more to SRY BS for mXYSRa/Enh13 to function. The additive effect of SOX9 and SRY via these BSs was verified using anin vitroassay. In conclusion, SOX9 BS and SRY BS function redundantlyin vivo, and at least one more functional sequence should exist in mXYSRa/Enh13.