Rce1, a novel transcriptional repressor, regulates cellulase gene expression by antagonizing the transactivator Xyr1 in <i>Trichoderma reesei</i>

Cao, Yanli; Zheng, Fanglin; Wang, Lei; Zhao, Guolei; Chen, Guanjun; Zhang, Weixin; Liu, Weifeng

doi:10.1111/mmi.13685

Cited by 91 publications

(87 citation statements)

References 47 publications

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“…In order to perform chromatin immunoprecipitation assays of TrSWI1, the pMDP tcu1 ‐ Trswi1 plasmid was first digested by Asc I and Not I to release the P tcu1 promoter. The P tcu1 promoter fused with the protein A coding sequence amplified with fusion extension PCR (Heckman and Pease, ) was then insert into the above digested pMDP tcu1 ‐ Trswi1 to obtain the pMDP tcu1 proA‐ Trswi1 plasmid, which was linearized with Hin dIII before being transformed into the QM9414, Δ xyr1 , OE xyr1 (Cao et al , ) and Δ Trsnf12 strains, respectively, to obtain the QM9414‐proA‐ Trswi1 , Δ xyr1 ‐proA‐ Trswi1 , OE xyr1 ‐proA‐ Trswi1 and Δ Trsnf12 ‐proA‐ Trswi1 strains.…”

Section: Methodsmentioning

confidence: 99%

Trichoderma reesei XYR1 recruits SWI/SNF to facilitate cellulase gene expression

Cao

Zheng

Zhang

et al. 2019

Molecular Microbiology

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Summary Cellulase gene expression in Trichoderma reesei is highly responsive to environmental cues and is under stringent regulation by multiple transcription factors. XYR1 (Xylanase regulator 1) has been identified as the most important transcriptional activator of cellulase/hemicellulase gene expression although the precise transactivating mechanism remains largely elusive. Here we show that the activation domain of XYR1 interacts with the T. reesei homolog of the TrSNF12 subunit of SWI/SNF complex. Deletion of Trsnf12 markedly impaired the induced cellulase gene expression. Individual loss of other SWI/SNF subunits including the catalytic subunit also severely compromised cellulase gene expression and interfered with loss of histone H4 in the cbh1 and eg1 promoters upon cellulose induction. In addition, we find that the SWI/SNF occupancy on cellulase gene promoters strictly required XYR1 and TrSNF12 but TrSNF12 was dispensable for the XYR1 binding to these promoters. These data suggest a model in which XYR1 recruits SWI/SNF through direct interactions with TrSNF12 to remodel chromatin at cellulase gene promoters, thereby activating cellulase gene expression to initiate the cellulolytic response in T. reesei.

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Section: Methodsmentioning

confidence: 99%

Trichoderma reesei XYR1 recruits SWI/SNF to facilitate cellulase gene expression

Cao

Zheng

Zhang

et al. 2019

Molecular Microbiology

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“…Several transcription factors (TFs) are now known for exerting modulation on the expression levels of holocellulases genes (Paula et al, 2018). In this sense, the regulatory network that encompasses cell wall deconstruction is under regulation of positive regulators, such as XYR1 (Stricker et al, 2006), ACEII (Aro et al, 2001), LAE1 (Seiboth et al, 2012), BglR (Nitta et al, 2012), VEL1 (Karimi et al, 2014), HAP 2/3/5 complex (Zeilinger et al, 2001) and other proteins which play a negative control in such process, as CRE1 (Portnoy et al, 2011), ACEI (Aro et al, 2003) and RCE1 (Cao et al, 2017). Additionally, it is already known that xylanases genes may also be targets of regulation by TFs, such as Xpp1 (Derntl et al, 2015b) and SxlR (Liu et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

CRZ1 regulator and calcium cooperatively modulate holocellulases gene expression in Trichoderma reesei QM6a

et al. 2020

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Trichoderma reesei is the main filamentous fungus used in industry to produce cellulases. Here we investigated the role of CRZ1 and Ca 2+ signaling in the fungus T. reesei QM6a concerning holocellulases production. For this, we first searched for potential CRZ1 binding sites in promoter regions of key genes coding holocellulases, as well as transcriptional regulators and sugar and calcium transporters. Using a nearly constructed T. reesei Dcrz1 strain, we demonstrated that most of the genes expected to be regulated by CRZ1 were affected in the mutant strain induced with sugarcane bagasse (SCB) and cellulose. In particular, our data demonstrate that Ca 2+ acts synergistically with CRZ1 to modulate gene expression, but also exerts CRZ1-independent regulatory role in gene expression in T. reesei, highlighting the role of the major regulator Ca 2+ on the signaling for holocellulases transcriptional control in the most part of cellulases genes here investigated. This work presents new evidence on the regulatory role of CRZ1 and Ca 2+ sensing in the regulation of cellulolytic enzymes in T. reesei, evidencing significant and previously unknown function of this Ca 2+ sensing system in the control key transcriptional regulators (XYR1 and CRE1) and on the expression of genes related to sugar and Ca 2+ transport.

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“…Immunoprecipitation was carried out through incubation of the anti-Myc antibodies or IgG (immunoglobulin G) with an aliquot of clarified cell lysates containing equal amounts of protein (2 mg) at 4°C for 5 h. Following this, the mixture was incubated with 40 l of protein A/G-beads which had been precoated with 1 mg/ml of BSA and 1 mg/ml of fish sperm DNA at 4°C for 4 h, followed by centrifugation to discard supernatant and extensive sequential washes as described previously (50). DNA was eluted with an elution buffer (100 mM Tris-HCl buffer [pH 7.8], plus 10 mM EDTA, 1% SDS, 10 mM NaHCO 3 , and 10 mM NaCl) at 65°C for 5 h. It was then recovered by proteinase K treatment of the pelleted samples at 45°C for 1 h and subjected to phenol-chloroform extraction and ethanol precipitation.…”

Section: Methodsmentioning

confidence: 99%

An Extracytoplasmic Function Sigma Factor Controls Cellulose Utilization by Regulating the Expression of an Outer Membrane Protein in Cytophaga hutchinsonii

Wang

Zhang

Zhou

et al. 2019

Appl Environ Microbiol

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The common soil cellulolytic bacterium known as Cytophaga hutchinsonii makes use of a unique but poorly understood strategy in order to utilize cellulose. While several genes have been identified as being an active part of the utilization of cellulose, the mechanism(s) by which C. hutchinsonii both (i) senses its environment and (ii) regulates the expression of those genes are not as yet known. In this study, we identified and characterized the gene CHU_3097 encoding an extracytoplasmic function (ECF) σ factor (σcel1), the disruption of which compromised C. hutchinsonii cellulose assimilation to a large degree. The σcel1 and its putative partner anti-σcel1, encoded by the CHU_3096 gene found immediately downstream from CHU_3097, copurified in vitro. The σcel1 was discovered to be associated with inner membrane when cells were cultured on glucose and yet was partially released from the membrane in response to cellulose. This release was found to occur on glucose when the anti-σcel1 was absent. Transcriptome analyses found a σcel1-regulated, cellulose-responsive gene regulon, within which an outer membrane protein encoding the gene CHU_1276, essential for cellulose utilization, was discovered to be significantly downregulated by CHU_3097 disruption. The expression of CHU_1276 almost fully restored cellulose utilization to the CHU_3097 mutant, demonstrating that CHU_1276 represents a critical regulatory target of σcel1. In this way, our study provided insights into the role of an ECF σ factor in coordinating the cellulolytic response of C. hutchinsonii. IMPORTANCE The common cellulolytic bacterium Cytophaga hutchinsonii uses a unique but poorly understood strategy in order to make use of cellulose. Throughout the process of cellulosic biomass breakdown, outer membrane proteins are thought to play key roles; this is evidenced by CHU_1276, which is required for the utilization of cellulose. However, the regulatory mechanism of its expression is not yet known. We found and characterized an extracytoplasmic function σ factor that is involved in coordinating the cellulolytic response of C. hutchinsonii by directly regulating the expression of CHU_1276. This study makes a contribution to our understanding of the regulatory mechanism used by C. hutchinsonii in order to adjust its genetic programs and so deal with novel environmental cues.

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Rce1, a novel transcriptional repressor, regulates cellulase gene expression by antagonizing the transactivator Xyr1 in Trichoderma reesei

Cited by 91 publications

References 47 publications

Trichoderma reesei XYR1 recruits SWI/SNF to facilitate cellulase gene expression

Trichoderma reesei XYR1 recruits SWI/SNF to facilitate cellulase gene expression

CRZ1 regulator and calcium cooperatively modulate holocellulases gene expression in Trichoderma reesei QM6a

An Extracytoplasmic Function Sigma Factor Controls Cellulose Utilization by Regulating the Expression of an Outer Membrane Protein in Cytophaga hutchinsonii

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