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Applied Microbiology and Biotechnology

, Volume 101, Issue 5, pp 2067–2078 | Cite as

A copper-responsive promoter replacement system to investigate gene functions in Trichoderma reesei: a case study in characterizing SAGA genes

  • Fanglin Zheng
  • Yanli Cao
  • Xinxing Lv
  • Lei Wang
  • Chunyan Li
  • Weixin Zhang
  • Guanjun Chen
  • Weifeng LiuEmail author
Applied genetics and molecular biotechnology

Abstract

Trichoderma reesei represents an important workhorse for industrial production of cellulases as well as other proteins. The molecular mechanism underlying the regulation of cellulase production as well as other physiological processes in T. reesei is still insufficiently understood. We constructed a P tcu1 -based promoter substitution cassette that allowed one-step replacement of the endogenous promoter for controlling the target gene expression with copper. We then showed that copper repression of the histone acetyltransferase gene gcn5 phenocopied the gcn5 deletion strain. Using the same strategy, we further characterized the function of another putative Spt-Ada-Gcn5 acetyltransferase (SAGA) complex subunit encoding gene, ada2, in T. reesei. Similar to the repression of gcn5, the addition of copper to the P tcu1 -ada2 strain not only drastically reduced the vegetative growth and conidiation in T. reesei but also severely compromised the induced cellulase gene expression. The developed strategy will thus be potentially useful to probe the biological function of the large fraction of T. reesei genes with unknown functions including those essential genes in the genome to expand its extraordinary biotechnological potential.

Keywords

tcu1 Gcn5 Ada2 Cellulase Trichoderma reesei 

Notes

Acknowledgements

This work is supported by grants from the National Natural Science Foundation of China (31270116, 31470162, and 31300059) and National Key Technology Support Program (2015BAD15B05). Zhang WX would also like to acknowledge the support from the Scientific Research Foundation for Excellent Young and Middle-Aged Scientists of Shandong Province of China (BS2013NJ021) and China Postdoctoral Science Foundation (2012M521325).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Research involving human participants and/or animals

This article does not contain any studies with human participants or animals performed by any of the authors.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Fanglin Zheng
    • 1
  • Yanli Cao
    • 1
  • Xinxing Lv
    • 1
  • Lei Wang
    • 1
  • Chunyan Li
    • 1
  • Weixin Zhang
    • 1
  • Guanjun Chen
    • 1
  • Weifeng Liu
    • 1
    Email author
  1. 1.State Key Laboratory of Microbial Technology, School of Life ScienceShandong UniversityJinanPeople’s Republic of China

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