Departamento de Ciencias Ecológicas, Facultad de Ciencias, Universidad de Chile, Santiago 7800003, Chile.
Instituto Andino Patagónico de Tecnologías Biológicas y Geoambientales (Consejo Nacional de Investigaciones Científicas y Técnicas), CONICET-UNCo, Universidad Nacional del Comahue, Bariloche 8400, Rio Negro, Argentina.
Int J Mol Sci. 2023 Mar 21;24(6):5930. doi: 10.3390/ijms24065930.
The UCD 67-385 genome harbors a 7873 bp cluster containing , and , encoding 2-desmethy-4-deoxygadusol synthase, O-methyl transferase, and ATP-grasp ligase, respectively, of the mycosporine glutaminol (MG) biosynthesis pathway. Homozygous deletion mutants of the entire cluster, single-gene mutants, and the Δ;Δ and Δ;Δ double-gene mutants did not produce mycosporines. However, Δ accumulated the intermediate 4-deoxygadusol. Heterologous expression of the and or , and cDNAs in led to 4-deoxygadusol or MG production, respectively. Genetic integration of the complete cluster into the genome of the non-mycosporine-producing CBS 6938 wild-type strain resulted in a transgenic strain (CBS 6938_MYC) that produced MG and mycosporine glutaminol glucoside. These results indicate the function of , and in the mycosporine biosynthesis pathway. The transcription factor gene mutants Δ, Δcyc8, and Δ showed upregulation, Δ and Δ showed downregulation, and Δ and Δ showed no effect on mycosporinogenesis in glucose-containing medium. Finally, comparative analysis of the cluster sequences in several strains and the four newly described species of the genus showed the phylogenetic relationship of the strains and their differentiation from the other species of the genus .
UCD67-385 基因组含有一个 7873bp 的簇，包含编码 2-去甲基-4-脱氮甘氨酸合成酶、O-甲基转移酶和 ATP 攫取连接酶的基因，分别为肌醇六磷酸谷氨酸（MG）生物合成途径的。整个簇的纯合缺失突变体、单基因突变体和Δ;Δ和Δ;Δ双基因突变体都不产生肌醇六磷酸谷氨酸。然而，Δ;Δ积累了中间产物 4-脱氮甘氨酸。将和或、和的 cDNA 在中异源表达分别导致 4-脱氮甘氨酸或 MG 的产生。将完整簇的遗传整合到非肌醇六磷酸谷氨酸产生的 CBS6938 野生型菌株的基因组中，导致产生 MG 和肌醇六磷酸谷氨酸葡萄糖苷的转基因菌株（CBS6938_MYC）。这些结果表明、和在肌醇六磷酸谷氨酸生物合成途径中的功能。在含有葡萄糖的培养基中，转录因子基因突变体Δ、Δcyc8 和 Δ显示上调，Δ和 Δ显示下调，而 Δ和 Δ对肌醇六磷酸谷氨酸生物合成没有影响。最后，对几个菌株中的簇序列进行比较分析和四个新描述的种属表明了菌株的系统发育关系及其与该属其他种属的分化。
