Involvement of BldC in the Formation of Physiologically Mature Sporangium in Actinoplanes missouriensis
Involvement of BldC in the Formation of Physiologically Mature Sporangium in Actinoplanes missouriensis
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BldC 参与密苏里游动放线菌生理成熟孢子囊的形成
DOI：
10.1128/jb.00189-22
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发表时间：
2022
影响因子：
3.2
通讯作者：
Ohnishi Yasuo
中科院分区：
文献类型：
--
作者：
Tezuka Takeaki;Nitta Shumpei;Ohnishi Yasuo
AmBldD是一个全局性转录调节因子，抑制密苏里游动放线菌孢子囊形成所需的几个基因的转录。在这里，我们的特点之一AmBldD的调节子：Vdd_1980，编码BldC，这是一个转录调节参与链霉菌的形态发育的直系同源物。通过高分辨率S1核酸酶图谱分析确定了bldCortholog的转录起始点，并在其5′端非翻译区发现了AmBldD盒。逆转录-定量PCR分析表明，bldC基因在孢子囊形成过程中被激活。结果表明，Δ bldCnull突变株（ΔbldC）形成的孢子囊形状正常，但孢子囊开裂缺陷，在诱导开裂条件下，Δ bldC突变株孢子囊释放的孢子数量比野生型菌株低2个数量级。RNA测序分析表明，BldC功能作为几个发育基因的转录激活因子，包括tcrA，它编码一个关键的转录激活因子，调节孢子囊形成，孢子囊开裂，孢子休眠。利用电泳迁移率变动分析（EMSA），我们发现重组BldC蛋白直接结合至少18个基因的上游区域，这些基因的转录在Δ bldC株中被下调。此外，使用DNase I足迹和EMSA，我们证明了BldC结合到含有AT丰富的基序的直接重复序列。因此，BldC是一个全局调节因子，它激活几个基因的转录，其中一些基因可能是孢子囊裂开所需的。重要的是，BldC是一个全局转录调节因子，在链霉菌的形态分化中充当“刹车”。BlDC样蛋白广泛分布于真细菌中，但其直系同源物尚未在链霉菌外进行研究。在这里，我们发现，BldC直系同源放线菌missouriensis是必不可少的孢子囊裂开，其调节子是不同的BldC调节子在委内瑞拉链霉菌，这表明BldC已演变成两个属的丝状放线菌之间的形态分化发挥不同的作用。
AmBldD is a global transcriptional regulator that represses the transcription of several genes required for sporangium formation in Actinoplanes missouriensis. Here, we characterized one of the AmBldD regulons:AMIS_1980, encoding an ortholog of BldC, which is a transcriptional regulator involved in the morphological development of Streptomyces. We determined the transcriptional start point of thebldCortholog by high-resolution S1 nuclease mapping and found an AmBldD box in its 5′-untranslated region. Reverse transcription-quantitative PCR analysis revealed that the transcription ofbldCis activated during sporangium formation. AbldCnull mutant (ΔbldC) strain formed normally shaped sporangia, but they exhibited defective sporangium dehiscence; under a dehiscence-inducing condition, the number of spores released from the sporangia of the ΔbldCstrain was 2 orders of magnitude lower than that from the sporangia of the wild-type strain. RNA sequencing analysis indicated that BldC functions as a transcriptional activator of several developmental genes, includingtcrA, which encodes a key transcriptional activator that regulates sporangium formation, sporangium dehiscence, and spore dormancy. Using electrophoretic mobility shift assay (EMSA), we showed that a recombinant BldC protein directly binds to upstream regions of at least 18 genes, the transcription of which is downregulated in the ΔbldCstrain. Furthermore, using DNase I footprinting and EMSA, we demonstrated that BldC binds to the direct repeat sequences containing an AT-rich motif. Thus, BldC is a global regulator that activates the transcription of several genes, some of which are likely to be required for sporangium dehiscence.IMPORTANCEBldC is a global transcriptional regulator that acts as a “brake” in the morphological differentiation of Streptomyces. BldC-like proteins are widely distributed throughout eubacteria, but their orthologs have not been studied outside streptomycetes. Here, we revealed that the BldC ortholog in Actinoplanes missouriensis is essential for sporangium dehiscence and that its regulon is different from the BldC regulon in Streptomyces venezuelae, suggesting that BldC has evolved to play different roles in morphological differentiation between the two genera of filamentous actinomycetes.
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影响因子：
11.3
作者：
McCormick JR;Flärdh K
通讯作者：
Flärdh K
影响因子：
3.2
作者：
Hunt, AC;Servín-González, L;Buttner, MJ
通讯作者：
Buttner, MJ
影响因子：
10.5
作者：
Willemse, Joost;Borst, Jan Willem;van Wezel, Gilles P.
通讯作者：
van Wezel, Gilles P.
密苏里游动放线菌孢子鞭毛的特征
影响因子：
4.4
作者：
K. Uchida;M. Jang;Y. Ohnishi;S. Horinouchi;M. Hayakawa;N. Fujita;S. Aizawa
通讯作者：
S. Aizawa
影响因子：
3.2
作者：
Hashiguchi, Yuichiro;Tezuka, Takeaki;Ohnishi, Yasuo
通讯作者：
Ohnishi, Yasuo