Overlapping coactivator function is required for transcriptional activation by the Candida glabrata Pdr1 transcription factor.

IF 3.3 3区 生物学 Q2 GENETICS & HEREDITY
Genetics Pub Date : 2024-09-04 DOI:10.1093/genetics/iyae115
Thomas P Conway, Lucia Simonicova, W Scott Moye-Rowley
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引用次数: 0

Abstract

Azole resistance in the pathogenic yeast Candida glabrata is a serious clinical complication and increasing in frequency. The majority of resistant organisms have been found to contain a substitution mutation in the Zn2Cys6 zinc cluster-containing transcription factor Pdr1. These mutations typically lead to this factor driving high, constitutive expression of target genes like the ATP-binding cassette transporter-encoding gene CDR1. Overexpression of Cdr1 is required for the observed elevated fluconazole resistance exhibited by strains containing one of these hyperactive PDR1 alleles. While the identity of hyperactive PDR1 alleles has been extensively documented, the mechanisms underlying how these gain-of-function (GOF) forms of Pdr1 lead to elevated target gene transcription are not well understood. We have used a tandem affinity purification-tagged form of Pdr1 to identify coactivator proteins that biochemically purify with the wild-type and 2 different GOF forms of Pdr1. Three coactivator proteins were found to associate with Pdr1: the SWI/SNF complex Snf2 chromatin remodeling protein and 2 different components of the SAGA complex, Spt7 and Ngg1. We found that deletion mutants lacking either SNF2 or SPT7 exhibited growth defects, even in the absence of fluconazole challenge. To overcome these issues, we employed a conditional degradation system to acutely deplete these coactivators and determined that loss of either coactivator complex, SWI/SNF or SAGA, caused defects in Pdr1-dependent transcription. A double degron strain that could be depleted for both SWI/SNF and SAGA exhibited a profound defect in PDR1 autoregulation, revealing that these complexes work together to ensure high-level Pdr1-dependent gene transcription.

念珠菌 Pdr1 转录因子的转录激活需要重叠的辅激活因子功能。
致病酵母光滑念珠菌的唑类抗药性是一种严重的临床并发症,而且越来越频繁。研究发现,大多数耐药菌体内含有 Zn2Cys6 锌簇转录因子 Pdr1 的替代突变。这些突变通常会导致该因子驱动 ATP 结合盒转运体编码基因 CDR1 等目标基因的高组成型表达。含有这些超活性 PDR1 等位基因之一的菌株所表现出的氟康唑抗药性升高需要 Cdr1 的过度表达。虽然超活性 PDR1 等位基因的特性已被广泛记录,但这些功能增益(GOF)形式的 Pdr1 如何导致靶基因转录升高的机制却不甚明了。我们使用串联亲和纯化(TAP)标记的 Pdr1 来鉴定与野生型和两种不同 GOF 形式的 Pdr1 一起进行生化纯化的辅激活因子蛋白。我们发现有三种辅激活蛋白与 Pdr1 有关联:SWI/SNF 复合物 Snf2 染色质重塑蛋白和 SAGA 复合物的两种不同成分 Spt7 和 Ngg1。我们发现,缺乏 SNF2 或 SPT7 的缺失突变体表现出生长缺陷,即使在没有氟康唑挑战的情况下也是如此。为了克服这些问题,我们采用了一种条件降解系统,以急性消耗这些辅激活因子,并确定 SWI/SNF 或 SAGA 辅激活因子复合物的缺失会导致 Pdr1 依赖性转录缺陷。同时缺失 SWI/SNF 和 SAGA 的双降解子菌株在 PDR1 自调节中表现出严重缺陷,这揭示了这些复合物共同确保高水平的 Pdr1 依赖性基因转录。
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来源期刊
Genetics
Genetics GENETICS & HEREDITY-
CiteScore
6.90
自引率
6.10%
发文量
177
审稿时长
1.5 months
期刊介绍: GENETICS is published by the Genetics Society of America, a scholarly society that seeks to deepen our understanding of the living world by advancing our understanding of genetics. Since 1916, GENETICS has published high-quality, original research presenting novel findings bearing on genetics and genomics. The journal publishes empirical studies of organisms ranging from microbes to humans, as well as theoretical work. While it has an illustrious history, GENETICS has changed along with the communities it serves: it is not your mentor''s journal. The editors make decisions quickly – in around 30 days – without sacrificing the excellence and scholarship for which the journal has long been known. GENETICS is a peer reviewed, peer-edited journal, with an international reach and increasing visibility and impact. All editorial decisions are made through collaboration of at least two editors who are practicing scientists. GENETICS is constantly innovating: expanded types of content include Reviews, Commentary (current issues of interest to geneticists), Perspectives (historical), Primers (to introduce primary literature into the classroom), Toolbox Reviews, plus YeastBook, FlyBook, and WormBook (coming spring 2016). For particularly time-sensitive results, we publish Communications. As part of our mission to serve our communities, we''ve published thematic collections, including Genomic Selection, Multiparental Populations, Mouse Collaborative Cross, and the Genetics of Sex.
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