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Abstract：
BACKGROUND: Epigenetic modifications of histones play important roles in the response of eukaryotic organisms to environmental stress. However, many histone acetyltransferases (HATs), which are responsible for histone acetylation, and their roles in mediating the tick response to cold stress have yet to be identified. In the present study, HATs were molecularly characterized and their associations with the cold response of the tick Haemaphysalis longicornis explored.
METHODS: HATs were characterized by using polymerase chain reaction (PCR) based on published genome sequences, followed by multiple bioinformatic analyses. The differential expression of genes in H. longicornis under different cold treatment conditions was evaluated using reverse transcription quantitative PCR (RT-qPCR). RNA interference was used to explore the association of HATs with the cold response of H. longicornis.
RESULTS: Two HAT genes were identified in H. longicornis (Hl), a GCN5-related N-acetyltransferase (henceforth HlGNAT) and a type B histone acetyltransferase (henceforth HlHAT-B), which are respectively 960 base pairs (bp) and 1239 bp in length. Bioinformatics analysis revealed that HlGNAT and HlHAT-B are unstable hydrophilic proteins characterized by the presence of the acetyltransferase 16 domain and Hat1_N domain, respectively. RT-qPCR revealed that the expression of HlGNAT and HlHAT-B decreased after 3 days of cold treatment, but gradually increased with a longer period of cold treatment. The mortality rate following knockdown of HlGNAT or HlHAT-B by RNA interference, which was confirmed by RT-qPCR, significantly increased (P < 0.05) when H. longicornis was treated at the lowest lethal temperature (- 14 °C) for 2 h.
CONCLUSIONS: The findings demonstrate that HATs may play a crucial role in the cold response of H. longicornis. Thus further research is warranted to explore the mechanisms underlying the epigenetic regulation of the cold response in ticks.
METHODS: HATs were characterized by using polymerase chain reaction (PCR) based on published genome sequences, followed by multiple bioinformatic analyses. The differential expression of genes in H. longicornis under different cold treatment conditions was evaluated using reverse transcription quantitative PCR (RT-qPCR). RNA interference was used to explore the association of HATs with the cold response of H. longicornis.
RESULTS: Two HAT genes were identified in H. longicornis (Hl), a GCN5-related N-acetyltransferase (henceforth HlGNAT) and a type B histone acetyltransferase (henceforth HlHAT-B), which are respectively 960 base pairs (bp) and 1239 bp in length. Bioinformatics analysis revealed that HlGNAT and HlHAT-B are unstable hydrophilic proteins characterized by the presence of the acetyltransferase 16 domain and Hat1_N domain, respectively. RT-qPCR revealed that the expression of HlGNAT and HlHAT-B decreased after 3 days of cold treatment, but gradually increased with a longer period of cold treatment. The mortality rate following knockdown of HlGNAT or HlHAT-B by RNA interference, which was confirmed by RT-qPCR, significantly increased (P < 0.05) when H. longicornis was treated at the lowest lethal temperature (- 14 °C) for 2 h.
CONCLUSIONS: The findings demonstrate that HATs may play a crucial role in the cold response of H. longicornis. Thus further research is warranted to explore the mechanisms underlying the epigenetic regulation of the cold response in ticks.
摘要：
背景：组蛋白的表观遗传修饰在真核生物对环境胁迫的响应中起重要作用。然而,许多组蛋白乙酰转移酶（HAT），负责组蛋白乙酰化，它们在介导蜱对冷应激的反应中的作用尚未确定。在本研究中,对HAT进行了分子表征，并探索了它们与壁虱长壁虱的冷反应的关系。
方法：根据已发表的基因组序列，使用聚合酶链反应（PCR）对HAT进行表征，其次是多个生物信息学分析。采用逆转录定量PCR(RT-qPCR)方法评价了不同冷处理条件下长尾H.longicornis基因的差异表达。RNA干扰用于探索HATs与长柄H.longicornis冷反应的关系。
结果：在H.longicornis（Hl）中鉴定出两个HAT基因，GCN5相关的N-乙酰转移酶（以下简称HlGNAT）和B型组蛋白乙酰转移酶（以下简称HlHAT-B），其长度分别为960个碱基对(bp)和1239bp。生物信息学分析表明,HlGNAT和HlHAT-B是不稳定的亲水性蛋白,其特征在于乙酰转移酶16结构域和Hat1_N结构域的存在,分别。RT-qPCR显示，冷处理3天后，HlGNAT和HlHAT-B的表达下降，但随着冷处理时间的延长而逐渐增加。通过RNA干扰敲除HlGNAT或HlHAT-B后的死亡率，通过RT-qPCR证实，当H.longicornis在最低致死温度（-14°C）下处理2h时，显着增加（P<0.05）。
结论：研究结果表明，HATs可能在H.longicornis的寒冷反应中起关键作用。因此，有必要进一步研究以探索壁虱冷反应的表观遗传调控机制。
方法：根据已发表的基因组序列，使用聚合酶链反应（PCR）对HAT进行表征，其次是多个生物信息学分析。采用逆转录定量PCR(RT-qPCR)方法评价了不同冷处理条件下长尾H.longicornis基因的差异表达。RNA干扰用于探索HATs与长柄H.longicornis冷反应的关系。
结果：在H.longicornis（Hl）中鉴定出两个HAT基因，GCN5相关的N-乙酰转移酶（以下简称HlGNAT）和B型组蛋白乙酰转移酶（以下简称HlHAT-B），其长度分别为960个碱基对(bp)和1239bp。生物信息学分析表明,HlGNAT和HlHAT-B是不稳定的亲水性蛋白,其特征在于乙酰转移酶16结构域和Hat1_N结构域的存在,分别。RT-qPCR显示，冷处理3天后，HlGNAT和HlHAT-B的表达下降，但随着冷处理时间的延长而逐渐增加。通过RNA干扰敲除HlGNAT或HlHAT-B后的死亡率，通过RT-qPCR证实，当H.longicornis在最低致死温度（-14°C）下处理2h时，显着增加（P<0.05）。
结论：研究结果表明，HATs可能在H.longicornis的寒冷反应中起关键作用。因此，有必要进一步研究以探索壁虱冷反应的表观遗传调控机制。
