ACE4, a Novel Transcriptional Activator Involved in the Regulation of Cellulase Genes during Growth on Cellulose.

Yumeng Chen, Aibo Lin, Pei Liu, Xingjia Fan, Chuan Wu, Ni Li, Liujing Wei, Wei Wang, Dongzhi Wei
Author Information
  1. Yumeng Chen: The State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China.
  2. Aibo Lin: The State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China.
  3. Pei Liu: The State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China.
  4. Xingjia Fan: The State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China.
  5. Chuan Wu: The State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China.
  6. Ni Li: The State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China.
  7. Liujing Wei: The State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China.
  8. Wei Wang: The State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China.
  9. Dongzhi Wei: The State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, China.

Abstract

The filamentous fungus Trichoderma reesei is a model strain for cellulase production. Cellulase gene expression in is controlled by multiple transcription factors. Here, we identified by comparative genomic screening a novel transcriptional activator, ACE4 (ctivator of ellulase xpression 4), that positively regulates cellulase gene expression on cellulose in . Disruption of the gene significantly decreased expression of four main cellulase genes and the essential cellulase transcription factor-encoding gene . Overexpression of increased cellulase production by approximately 22% compared to that in the parental strain. Further investigations using electrophoretic mobility shift assays, DNase I footprinting assays, and chromatin immunoprecipitation assays indicated that ACE4 directly binds to the promoter of cellulase genes by recognizing the two adjacent 5'-GGCC-3' sequences. Additionally, ACE4 directly binds to the promoter of and, in turn, regulates the expression of ACE3 to facilitate cellulase production. Collectively, these results demonstrate an important role for ACE4 in regulating cellulase gene expression, which will contribute to understanding the mechanism underlying cellulase expression in . is commonly utilized in industry to produce cellulases, enzymes that degrade lignocellulosic biomass for the production of bioethanol and bio-based products. is capable of rapidly initiating the biosynthesis of cellulases in the presence of cellulose, which has made it useful as a model fungus for studying gene expression in eukaryotes. Cellulase gene expression is controlled through multiple transcription factors at the transcriptional level. However, the molecular mechanisms by which transcription is controlled remain unclear. In the present study, we identified a novel transcription factor, ACE4, which regulates cellulase expression on cellulose by binding to the promoters of cellulase genes and the cellulase activator gene . Our study not only expands the general functional understanding of the novel transcription factor ACE4 but also provides evidence for the regulatory mechanism mediating gene expression in .

Keywords

References

  1. Biotechnol Biofuels. 2018 Mar 1;11:54 [PMID: 29507606]
  2. Microb Cell Fact. 2016 Jun 10;15(1):106 [PMID: 27287427]
  3. Biotechnol Biofuels. 2014 Jan 28;7(1):14 [PMID: 24472375]
  4. Biotechnol Biofuels. 2009 Sep 01;2:19 [PMID: 19723296]
  5. Mol Biol Evol. 2013 Dec;30(12):2725-9 [PMID: 24132122]
  6. Eukaryot Cell. 2011 Feb;10(2):262-71 [PMID: 21169417]
  7. J Biotechnol. 2010 Jan 1;145(1):30-7 [PMID: 19861137]
  8. Fungal Genet Biol. 2008 Apr;45(4):436-45 [PMID: 17920314]
  9. Biotechnol Biofuels. 2015 Apr 14;8:67 [PMID: 25926888]
  10. Biotechnol Biofuels. 2017 Sep 7;10:209 [PMID: 28912831]
  11. Biotechnol Adv. 2019 Jul - Aug;37(4):519-529 [PMID: 30576717]
  12. Fungal Biol Biotechnol. 2018 Aug 20;5:15 [PMID: 30151221]
  13. Biotechnol Biofuels. 2017 Feb 2;10:30 [PMID: 28184245]
  14. Annu Rev Microbiol. 2013;67:477-98 [PMID: 23808333]
  15. Mol Genet Genomics. 2003 Oct;270(1):46-55 [PMID: 12905071]
  16. Biotechnol Biofuels. 2017 Jun 12;10:152 [PMID: 28616076]
  17. Mol Microbiol. 2016 May;100(3):560-75 [PMID: 27109892]
  18. Plasmid. 2012 Jan;67(1):67-71 [PMID: 22056690]
  19. J Bacteriol. 2012 Oct;194(19):5237-44 [PMID: 22821977]
  20. J Biol Chem. 2000 Feb 25;275(8):5817-25 [PMID: 10681571]
  21. Biotechnol Bioeng. 2020 Jun;117(6):1747-1760 [PMID: 32124970]
  22. Sci Rep. 2016 Feb 09;6:20761 [PMID: 26857594]
  23. Biotechnol Biofuels. 2013 Sep 09;6(1):127 [PMID: 24016404]
  24. Eukaryot Cell. 2006 Dec;5(12):2128-37 [PMID: 17056741]
  25. J Ind Microbiol Biotechnol. 2013 Jun;40(6):633-41 [PMID: 23467998]
  26. Biotechnol Biofuels. 2020 Apr 1;13:62 [PMID: 32266008]
  27. Nat Protoc. 2008;3(10):1671-8 [PMID: 18833205]
  28. Nucleic Acids Res. 2012 Jan;40(Database issue):D290-301 [PMID: 22127870]
  29. Microbiology (Reading). 2012 Jan;158(Pt 1):58-68 [PMID: 21998163]
  30. Mol Gen Genet. 1996 Jun 24;251(4):451-60 [PMID: 8709949]
  31. Biotechnol Adv. 2019 Nov 1;37(6):107347 [PMID: 30771467]
  32. J Biol Chem. 2019 Nov 29;294(48):18435-18450 [PMID: 31501242]
  33. J Biol Chem. 2001 Jun 29;276(26):24309-14 [PMID: 11304525]
  34. Biotechnol Biofuels. 2019 Oct 8;12:238 [PMID: 31624500]
  35. Biotechnol Biofuels. 2019 Feb 19;12:36 [PMID: 30820246]
  36. Mol Microbiol. 2017 Jul;105(1):65-83 [PMID: 28378498]
  37. Front Microbiol. 2016 Feb 16;7:175 [PMID: 26909077]
  38. Trends Biochem Sci. 2016 Jul;41(7):633-645 [PMID: 27211037]
  39. PLoS Genet. 2020 Sep 2;16(9):e1008979 [PMID: 32877410]
  40. Microb Cell Fact. 2019 May 10;18(1):81 [PMID: 31077201]

MeSH Term

Cellulase
Cellulose
Cellulose 1,4-beta-Cellobiosidase
Endo-1,4-beta Xylanases
Gene Expression Regulation, Fungal
Trans-Activators
Trichoderma

Chemicals

Trans-Activators
Cellulose
Cellulase
Endo-1,4-beta Xylanases
Cellulose 1,4-beta-Cellobiosidase

Word Cloud

Created with Highcharts 10.0.0cellulasegeneexpressionACE4transcriptionproductionCellulasecontrollednovelregulatescellulosegenesassaysfactorfungusTrichodermareeseimodelstrainmultiplefactorsidentifiedtranscriptionalactivatordirectlybindspromoterunderstandingmechanismcellulasesstudyfilamentouscomparativegenomicscreeningctivatorellulasexpression4positivelyDisruptionsignificantlydecreasedfourmainessentialfactor-encodingOverexpressionincreasedapproximately22%comparedparentalinvestigationsusingelectrophoreticmobilityshiftDNasefootprintingchromatinimmunoprecipitationindicatedrecognizingtwoadjacent5'-GGCC-3'sequencesAdditionallyturnACE3facilitateCollectivelyresultsdemonstrateimportantroleregulatingwillcontributeunderlyingcommonlyutilizedindustryproduceenzymesdegradelignocellulosicbiomassbioethanolbio-basedproductscapablerapidlyinitiatingbiosynthesispresencemadeusefulstudyingeukaryoteslevelHowevermolecularmechanismsremainunclearpresentbindingpromotersexpandsgeneralfunctionalalsoprovidesevidenceregulatorymediatingNovelTranscriptionalActivatorInvolvedRegulationGenesGrowthCelluloseZnII2Cys6protein

Similar Articles

Cited By