Dynamic changes in the extracellular proteome caused by absence of a pleiotropic regulator AdpA in Streptomyces griseus
Dynamic changes in the extracellular proteome caused by absence of a pleiotropic regulator AdpA in Streptomyces griseus
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DOI：
10.1111/j.1365-2958.2009.06814.x
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发表时间：
2009-09-01
影响因子：
3.6
通讯作者：
Horinouchi, Sueharu
中科院分区：
文献类型：
--
作者：
Akanuma, Genki;Hara, Hirofumi;Horinouchi, Sueharu
在灰色链霉菌中，A因子(2-isocapryloyl-3R-hydroxymethyl-gamma-butyrolactone)通过诱导多效性转录调节因子AdpA来触发形态发育和次生代谢。对野生型和Delta adpA菌株的胞外蛋白质组分析表明，AdpA依赖于AdpA的方式产生38种分泌蛋白质，其中包括许多分解代谢酶，如蛋白酶、糖基水解酶和酯酶。转录组分析表明，几乎所有这些依赖AdpA的分泌蛋白都在转录水平上受到调控。体外AdpA结合分析和转录起始点的确定导致11个启动子被确定为AdpA的新靶点。活力染色显示，在指数生长期有部分菌丝裂解，这可以解释在野生型和Delta adpA菌株的培养液中分别检测到3种和23种细胞质蛋白。在野生型菌株中，由于培养基中有较高的蛋白酶活性，从死亡细胞中渗出的细胞质蛋白似乎被降解并被重复使用，以供进一步生长。许多依赖AdpA的(即A因子可诱导的)分泌分解代谢酶的存在，可能参与死亡细胞渗漏物质的同化，再次强调了A因子在灰链霉菌形态分化中的重要性。
P>In Streptomyces griseus, A-factor (2-isocapryloyl-3R-hydroxymethyl-gamma-butyrolactone) triggers morphological development and secondary metabolism by inducing a pleiotropic transcriptional regulator AdpA. Extracellular proteome analysis of the wild-type and Delta adpA strains grown to the end of the exponential phase in liquid minimal medium revealed that 38 secreted proteins, including many catabolic enzymes, such as protease, glycosyl hydrolase and esterase, were produced in an AdpA-dependent manner. Transcriptome analysis showed that almost all of these AdpA-dependent secreted proteins were regulated at the transcriptional level. In vitro AdpA-binding assays and determination of transcriptional start sites led to identification of 11 promoters as novel targets of AdpA. Viability staining revealed that some hyphae lysed during the exponential growth phase, which could explain the detection of 3 and 23 cytoplasmic proteins in the culture media of the wild-type and Delta adpA strains respectively. In the wild-type strain, due to high protease activity in the culture medium, cytoplasmic proteins that leaked from dead cells seemed to be degraded and reused for the further growth. The existence of many AdpA-dependent (i.e. A-factor-inducible) secreted catabolic enzymes, which are likely involved in the assimilation of material that leaked from dead cells, reemphasizes the importance of A-factor in the morphological differentiation of S. griseus.