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盘基网柄菌磷酸二酯酶抑制剂阴性突变体中环状核苷酸磷酸二酯酶基因表达的调控

Regulation of expression of the cyclic nucleotide phosphodiesterase gene in phosphodiesterase inhibitor-negative mutants of Dictyostelium discoideum.

作者信息

Adames N R, Coukell M B, Wu L

机构信息

Department of Biology, York University, North York, ON Canada.

出版信息

Biochem Cell Biol. 1994 May-Jun;72(5-6):233-8. doi: 10.1139/o94-033.

Abstract

During early development of Dictyostelium discoideum, the enzyme cyclic nucleotide phosphodiesterase (PD) is produced at a low rate during the period its specific inhibitor (PDI) is being synthesized. In addition, PD gene expression is derepressed in the aggregation-deficient (Agg-), Pdi- mutant HC35. These observations suggest that the PDI might function to regulate PD gene expression, as well as modulate its activity. To explore this idea further, five new Agg-, Pdi- mutants were isolated and analyzed. All of the mutants produced high PD activity and overexpressed PD mRNA; four exhibited elevated levels of the 2400-nucleotide aggregation transcript and one overproduced the 1900-nucleotide vegetative transcript. In contrast, PD transcripts were not elevated in two Agg-, Pdi+ mutants. To determine if PDI production regulates PD expression, HC35 cells were transformed with plasmids carrying the PDI structural gene under the control of either the vegetative or aggregative PD promoter. Neither expression of PDI by the transformants nor addition of partially purified PDI to HC35 cells affected PD transcription. These results suggest that PD overexpression in the Pdi- mutants is not a direct consequence of the inability of these cells to produce inhibitor.

摘要

在盘基网柄菌的早期发育过程中,环核苷酸磷酸二酯酶(PD)在其特异性抑制剂(PDI)合成期间的产生速率较低。此外,在聚集缺陷型(Agg-)、Pdi-突变体HC35中,PD基因表达去抑制。这些观察结果表明,PDI可能在调节PD基因表达以及调节其活性方面发挥作用。为了进一步探究这一想法,分离并分析了五个新的Agg-、Pdi-突变体。所有突变体均产生高PD活性并过度表达PD mRNA;四个突变体表现出2400个核苷酸的聚集转录本水平升高,一个过度产生1900个核苷酸的营养转录本。相比之下,在两个Agg-、Pdi+突变体中,PD转录本没有升高。为了确定PDI的产生是否调节PD表达,用携带在营养型或聚集型PD启动子控制下的PDI结构基因的质粒转化HC35细胞。转化体对PDI的表达或向HC35细胞中添加部分纯化的PDI均不影响PD转录。这些结果表明,Pdi-突变体中PD的过度表达不是这些细胞无法产生抑制剂的直接后果。

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