HAP-Like CCAAT-Binding Complexes in Filamentous Fungi: Implications for Biotechnology

Brakhage, Axel A.; Andrianopoulos, Alex; Kato, Masashi; Steidl, Stefan; Davis, Meryl A.; Tsukagoshi, Norihiro; Hynes, Michael J.

doi:10.1006/fgbi.1999.1136

Cited by 104 publications

(62 citation statements)

References 53 publications

(70 reference statements)

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“…The presence of XLNR consensus binding sites in the promoter suggests that there is an induction process similar to that described for A. niger (52). Furthermore, the CCAAT box indicates that HAP complexmediated regulation occurs, as described for other xylanases (4,56). In A. nidulans two xylanases show an opposite pattern of expression with respect to ambient pH which is PACC mediated (36).…”

Section: Discussionmentioning

confidence: 83%

xylP Promoter-Based Expression System and Its Use for Antisense Downregulation of the Penicillium chrysogenum Nitrogen Regulator NRE

Zadra¹,

Abt²,

Parson

et al. 2000

Appl Environ Microbiol

117

124

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A highly inducible fungal promoter derived from the Penicillium chrysogenum endoxylanase (xylP) gene is described. Northern analysis and the use of a ␤-glucuronidase (uidA) reporter gene strategy showed that xylP expression is transcriptionally regulated. Xylan and xylose are efficient inducers, whereas glucose strongly represses the promoter activity. Comparison of the same expression construct as a single copy at the niaD locus in P. chrysogenum and at the argB locus in Aspergillus nidulans demonstrated that the xylP promoter is regulated similarly in these two species but that the level of expression is about 80 times higher in the Aspergillus species. The xylP promoter was found to be 65-fold more efficient than the isopenicillin-N-synthetase (pcbC) promoter in Penicillium and 23-fold more efficient than the nitrate reductase (niaD) promoter in Aspergillus under induced conditions. Furthermore, the xylP promoter was used for controllable antisense RNA synthesis of the nre-encoded putative major nitrogen regulator of P. chrysogenum. This approach led to inducible downregulation of the steady-state mRNA level of nre and consequently to transcriptional repression of the genes responsible for nitrate assimilation. In addition, transcription of nreB, which encodes a negative-acting nitrogen regulatory GATA factor of Penicillium, was found to be subject to regulation by NRE. Our data are the first direct evidence that nre indeed encodes an activator in the nitrogen regulatory circuit in Penicillium and indicate that cross regulation of the controlling factors occurs.

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Section: Discussionmentioning

confidence: 83%

xylP Promoter-Based Expression System and Its Use for Antisense Downregulation of the Penicillium chrysogenum Nitrogen Regulator NRE

Zadra¹,

Abt²,

Parson

et al. 2000

Appl Environ Microbiol

117

124

View full text Add to dashboard Cite

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“…CCAAT sequences have been found to be a binding site for the AnCF transcription factor (comprising the HapB, HapC, and HapE subunits) which affects, both negatively and positively, the expression of a diverse range of genes in A. nidulans (7,34,41). The expression of the acuF-lacZ fusion was found to be reduced by approximately 60% in a hapC deletion mutant background, but induction by proline or acetate was not affected (Fig.…”

Section: Resultsmentioning

confidence: 98%

Regulation of theacuFGene, Encoding Phosphoenolpyruvate Carboxykinase in the Filamentous FungusAspergillus nidulans

2002

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Phosphoenolpyruvate carboxykinase (PEPCK) is a key enzyme required for gluconeogenesis when microorganisms grow on carbon sources metabolized via the tricarboxylic acid (TCA) cycle. Aspergillus nidulans acuF mutants isolated by their inability to use acetate as a carbon source specifically lack PEPCK. The acuF gene has been cloned and shown to encode a protein with high similarity to PEPCK from bacteria, plants, and fungi. The regulation of acuF expression has been studied by Northern blotting and by the construction of lacZ fusion reporters. Induction by acetate is abolished in mutants unable to metabolize acetate via the TCA cycle, and induction by amino acids metabolized via 2-oxoglutarate is lost in mutants unable to form 2-oxoglutarate. Induction by acetate and proline is not additive, consistent with a single mechanism of induction. Malate and succinate result in induction, and it is proposed that PEPCK is controlled by a novel mechanism of induction by a TCA cycle intermediate or derivative, thereby allowing gluconeogenesis to occur during growth on any carbon source metabolized via the TCA cycle. It has been shown that the facB gene, which mediates acetate induction of enzymes specifically required for acetate utilization, is not directly involved in PEPCK induction. This is in contrast to Saccharomyces cerevisiae, where Cat8p and Sip4p, homologs of FacB, regulate PEPCK as well as the expression of other genes necessary for growth on nonfermentable carbon sources in response to the carbon source present. This difference in the control of gluconeogenesis reflects the ability of A. nidulans and other filamentous fungi to use a wide variety of carbon sources in comparison with S. cerevisiae. The acuF gene was also found to be subject to activation by the CCAAT binding protein AnCF, a protein homologous to the S. cerevisiae Hap complex and the mammalian NFY complex.

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“…Moreover, the Hap32 and Hap5 proteins also could heterodimerize through their histone-fold domains (as reviewed in Ref. 82), and in association with Hap2 could constitute the core HAP regulatory complex. Such a mechanism for complex formation has previously been reported for the S. cerevisiae Hap2-Hap3-Hap5 complex (29).…”

Section: Cap2-hap Regulatory Networkmentioning

confidence: 99%

Cap2-HAP Complex Is a Critical Transcriptional Regulator That Has Dual but Contrasting Roles in Regulation of Iron Homeostasis in Candida albicans

Singh¹,

Prasad²,

Sinha³

et al. 2011

Journal of Biological Chemistry

100

211

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Iron homeostasis is highly regulated in organisms across evolutionary time scale as iron is essential for various cellular processes. In a computational screen, we identified the Yap/bZIP domain family in Candida clade genomes. Cap2/Hap43 is essential for C. albicans growth under iron-deprivation conditions and for virulence in mouse. Cap2 has an amino-terminal bipartite domain comprising a fungal-specific Hap4-like domain and a bZIP domain. Our mutational analyses showed that both the bZIP and Hap4-like domains perform critical and independent functions for growth under iron-deprivation conditions. Transcriptome analysis conducted under iron-deprivation conditions identified about 16% of the C. albicans ORFs that were differentially regulated in a Cap2-dependent manner. Microarray data also suggested that Cap2 is required to mobilize iron through multiple mechanisms; chiefly by activation of genes in three iron uptake pathways and repression of iron utilizing and iron storage genes. The expression of HAP2, HAP32, and HAP5, core components of the HAP regulatory complex was induced in a Cap2-dependent manner indicating a feed-forward loop. In a feed-back loop, Cap2 repressed the expression of Sfu1, a negative regulator of iron uptake genes. Cap2 was coimmunoprecipitated with Hap5 from cell extracts prepared from iron-deprivation conditions indicating an in vivo association. ChIP assays demonstrated Hap32-dependent recruitment of Hap5 to the promoters of FRP1 (Cap2-induced) and ACO1 (Cap2-repressed). Together our data indicates that the Cap2-HAP complex functions both as a positive and a negative regulator to maintain iron homeostasis in C. albicans.

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HAP-Like CCAAT-Binding Complexes in Filamentous Fungi: Implications for Biotechnology

Cited by 104 publications

References 53 publications

xylP Promoter-Based Expression System and Its Use for Antisense Downregulation of the Penicillium chrysogenum Nitrogen Regulator NRE

xylP Promoter-Based Expression System and Its Use for Antisense Downregulation of the Penicillium chrysogenum Nitrogen Regulator NRE

Regulation of theacuFGene, Encoding Phosphoenolpyruvate Carboxykinase in the Filamentous FungusAspergillus nidulans

Cap2-HAP Complex Is a Critical Transcriptional Regulator That Has Dual but Contrasting Roles in Regulation of Iron Homeostasis in Candida albicans

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