Abstract
Blakeslea trispora is known for its potential to produce an excess of carotenoids in mixed cultures of strains of opposite sex. The biosynthesis of β-carotene in B. trispora is activated not only by sex hormone trisporic acid but also by light, especially blue light. In fungi, the most intensively investigated blue-light reception proteins are WC-1 and WC-2, and the two proteins form a transcription factor complex which is called WCC by their PAS domains. Notably, multiple genes similar to wc-1 and wc-2 have been identified and characterized in Phycomyces, Mucor, and Rhizopus. Here we report that there are four members of wc-2-like gene family in B. trispora genome: Btwc-2a, Btwc-2b, Btwc-2c, and Btwc-2d. When the mycelia were exposed to blue light, their transcription levels are regulated differentially. Except for BtWC-2b, which only has a PAS domain, the other three proteins contain both a PAS domain and a ZnF domain. BtWC-2a interacts with either BtWC-1a or BtWC-1c to form different photoreceptor complexes in yeast two-hybrid assays, which is the unique situation not yet described in other fungi. In addition, the protein–protein docking analysis by the predicted 3D structures showed that the two complexes are structurally different. These results suggested that WC proteins of B. trispora are still involved in light regulation by forming WCC and the regulation mechanism of the photobiology appears to be more complex.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Code availability
Not applicable.
References
Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402. https://doi.org/10.1093/nar/25.17.3389
Avalos J, Estrada AF (2010) Regulation by light in Fusarium. Fungal Genet Biol 47:930–938. https://doi.org/10.1016/j.fgb.2010.05.001
Ballario P, Vittorioso P, Magrelli A, Talora C, Cabibbo A, Macino G (1996) White collar-1, a central regulator of blue light responses in Neurospora, is a zinc finger protein. EMBO J 15:1650–1657
Bergman K, Eslava AP, Cerda-Olmedo E (1973) Mutants of Phycomyces with abnormal phototropism. Mol Gen Genet 123:1–16. https://doi.org/10.1007/BF00282984
Castrillo M, Avalos J (2015) The flavoproteins CryD and VvdA cooperate with the white collar protein WcoA in the control of photocarotenogenesis in Fusarium fujikuroi. PLoS ONE 10:e0119785. https://doi.org/10.1371/journal.pone.0119785
Cerda-Olmedo E (2001) Phycomyces and the biology of light and color. FEMS Microbiol Rev 25:503–512. https://doi.org/10.1111/j.1574-6976.2001.tb00588.x
Chen CH, Ringelberg CS, Gross RH, Dunlap JC, Loros JJ (2009) Genome-wide analysis of light-inducible responses reveals hierarchical light signalling in Neurospora. EMBO J 28:1029–1042. https://doi.org/10.1038/emboj.2009.54
Cheng P, Yang Y, Gardner KH, Liu Y (2002) PAS domain-mediated WC-1/WC-2 interaction is essential for maintaining the steady-state level of WC-1 and the function of both proteins in circadian clock and light responses of Neurospora. Mol Cell Biol 22:517–524. https://doi.org/10.1128/mcb.22.2.517-524.2002
Corrochano LM (2019) Light in the fungal world: from photoreception to gene transcription and beyond. Annu Rev Genet 53:149–170. https://doi.org/10.1146/annurev-genet-120417-031415
Corrochano LM, Garre V (2010) Photobiology in the zygomycota: multiple photoreceptor genes for complex responses to light. Fungal Genet Biol 47:893–899. https://doi.org/10.1016/j.fgb.2010.04.007
Corrochano LM et al (2016) Expansion of signal transduction pathways in fungi by extensive genome duplication. Curr Biol 26:1577–1584. https://doi.org/10.1016/j.cub.2016.04.038
Dasgupta A, Fuller KK, Dunlap JC, Loros JJ (2016) Seeing the world differently: variability in the photosensory mechanisms of two model fungi. Environ Microbiol 18:5–20. https://doi.org/10.1111/1462-2920.13055
Guex N, Peitsch MC, Schwede T (2009) Automated comparative protein structure modeling with SWISS-MODEL and Swiss-PdbViewer: a historical perspective. Electrophoresis 30(Suppl 1):S162-173. https://doi.org/10.1002/elps.200900140
Hu B, Jin J, Guo AY, Zhang H, Luo J, Gao G (2015) GSDS 2.0: an upgraded gene feature visualization server. Bioinformatics 31:1296–1297. https://doi.org/10.1093/bioinformatics/btu817
Idnurm A, Rodriguez-Romero J, Corrochano LM, Sanz C, Iturriaga EA, Eslava AP, Heitman J (2006) The Phycomyces madA gene encodes a blue-light photoreceptor for phototropism and other light responses. Proc Natl Acad Sci U S A 103:4546–4551. https://doi.org/10.1073/pnas.0600633103
Krissinel E, Henrick K (2007) Inference of macromolecular assemblies from crystalline state. J Mol Biol 372:774–797. https://doi.org/10.1016/j.jmb.2007.05.022
Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874. https://doi.org/10.1093/molbev/msw054
Kuratani M, Tanaka K, Terashima K, Muraguchi H, Nakazawa T, Nakahori K, Kamada T (2010) The dst2 gene essential for photomorphogenesis of Coprinopsis cinerea encodes a protein with a putative FAD-binding-4 domain. Fungal Genet Biol 47:152–158. https://doi.org/10.1016/j.fgb.2009.10.006
Linden H, Macino G (1997) White collar 2, a partner in blue-light signal transduction, controlling expression of light-regulated genes in Neurospora crassa. EMBO J 16:98–109. https://doi.org/10.1093/emboj/16.1.98
Lopez-Nieto MJ et al (2004) Biotechnological lycopene production by mated fermentation of Blakeslea trispora. Appl Microbiol Biotechnol 66:153–159. https://doi.org/10.1007/s00253-004-1669-4
Luo W, Xue C, Zhao Y, Zhang H, Rao Z, Yu X (2020) Blakeslea trispora photoreceptors: identification and functional analysis Appl Environ Microbiol 86 https://doi.org/10.1128/AEM.02962-19
Ma LJ et al (2009) Genomic analysis of the basal lineage fungus Rhizopus oryzae reveals a whole-genome duplication. PLoS Genet 5:e1000549. https://doi.org/10.1371/journal.pgen.1000549
Nagy G, Szebenyi C, Csernetics A, Vaz AG, Toth EJ, Vagvolgyi C, Papp T (2017) Development of a plasmid free CRISPR-Cas9 system for the genetic modification of Mucor circinelloides. Sci Rep 7:16800. https://doi.org/10.1038/s41598-017-17118-2
Nagy G et al (2019) CRISPR-Cas9-mediated disruption of the HMG-CoA reductase genes of Mucor circinelloides and subcellular localization of the encoded enzymes. Fungal Genet Biol 129:30–39. https://doi.org/10.1016/j.fgb.2019.04.008
Navarro E, Penaranda A, Hansberg W, Torres-Martinez S, Garre V (2013) A white collar 1-like protein mediates opposite regulatory functions in Mucor circinelloides. Fungal Genet Biol 52:42–52. https://doi.org/10.1016/j.fgb.2012.12.003
Pierce BG, Wiehe K, Hwang H, Kim BH, Vreven T, Weng Z (2014) ZDOCK server: interactive docking prediction of protein-protein complexes and symmetric multimers. Bioinformatics 30:1771–1773. https://doi.org/10.1093/bioinformatics/btu097
Polaino S et al (2017) A Ras GTPase associated protein is involved in the phototropic and circadian photobiology responses in fungi Sci Rep 7:44790. https://doi.org/10.1038/srep44790
Pontius J, Richelle J, Wodak SJ (1996) Deviations from standard atomic volumes as a quality measure for protein crystal structures. J Mol Biol 264:121–136. https://doi.org/10.1006/jmbi.1996.0628
Purschwitz J et al (2008) Functional and physical interaction of blue- and red-light sensors in Aspergillus nidulans. Curr Biol 18:255–259. https://doi.org/10.1016/j.cub.2008.01.061
Quiles-Rosillo MD, Ruiz-Vazquez RM, Torres-Martinez S, Garre V (2005) Light induction of the carotenoid biosynthesis pathway in Blakeslea trispora. Fungal Genet Biol 42:141–153. https://doi.org/10.1016/j.fgb.2004.10.008
Rodriguez-Romero J, Corrochano LM (2006) Regulation by blue light and heat shock of gene transcription in the fungus Phycomyces: proteins required for photoinduction and mechanism for adaptation to light. Mol Microbiol 61:1049–1059. https://doi.org/10.1111/j.1365-2958.2006.05293.x
Rodriguez-Saiz M, Paz B, De La Fuente JL, Lopez-Nieto MJ, Cabri W, Barredo JL (2004) Blakeslea trispora genes for carotene biosynthesis. Appl Environ Microbiol 70:5589–5594. https://doi.org/10.1128/AEM.70.9.5589-5594.2004
Sancar C, Ha N, Yilmaz R, Tesorero R, Fisher T, Brunner M, Sancar G (2015) Combinatorial control of light induced chromatin remodeling and gene activation in Neurospora. PLoS Genet 11:e1005105. https://doi.org/10.1371/journal.pgen.1005105
Sanz C, Rodriguez-Romero J, Idnurm A, Christie JM, Heitman J, Corrochano LM, Eslava AP (2009) Phycomyces MADB interacts with MADA to form the primary photoreceptor complex for fungal phototropism. Proc Natl Acad Sci U S A 106:7095–7100. https://doi.org/10.1073/pnas.0900879106
Schmidle A (1951) Die Tagesperiodizität der asexuellen Reproduktion von Pilobolus sphaerosporus. Arch Microbiol 16 https://doi.org/10.1007/BF00408955
Sevgili A, Erkmen O (2019) Improved lycopene production from different substrates by mated fermentation of Blakeslea Trispora. Foods 8 10.3390/foods8040120
Shakya VPS, Idnurm A (2017) The inhibition of mating in Phycomyces blakesleeanus by light is dependent on the MadA-MadB complex that acts in a sex-specific manner. Fungal Genet Biol 101:20–30. https://doi.org/10.1016/j.fgb.2017.02.005
Silva F, Navarro E, Penaranda A, Murcia-Flores L, Torres-Martinez S, Garre V (2008) A RING-finger protein regulates carotenogenesis via proteolysis-independent ubiquitylation of a white collar-1-like activator. Mol Microbiol 70:1026–1036. https://doi.org/10.1111/j.1365-2958.2008.06470.x
Silva F, Torres-Martinez S, Garre V (2006) Distinct white collar-1 genes control specific light responses in Mucor circinelloides. Mol Microbiol 61:1023–1037. https://doi.org/10.1111/j.1365-2958.2006.05291.x
Smith KM et al (2010) Transcription factors in light and circadian clock signaling networks revealed by genomewide mapping of direct targets for neurospora white collar complex. Eukaryot Cell 9:1549–1556. https://doi.org/10.1128/EC.00154-10
Terashima K, Yuki K, Muraguchi H, Akiyama M, Kamada T (2005) The dst1 gene involved in mushroom photomorphogenesis of Coprinus cinereus encodes a putative photoreceptor for blue light. Genetics 171:101–108. https://doi.org/10.1534/genetics.104.040048
Tisch D, Schmoll M (2010) Light regulation of metabolic pathways in fungi. Appl Microbiol Biotechnol 85:1259–1277. https://doi.org/10.1007/s00253-009-2320-1
Torres-Martinez S, Ruiz-Vazquez RM, Garre V, Lopez-Garcia S, Navarro E, Vila A (2012) Molecular tools for carotenogenesis analysis in the zygomycete Mucor circinelloides. Methods Mol Biol 898:85–107. https://doi.org/10.1007/978-1-61779-918-1_5
Wang B, Kettenbach AN, Zhou X, Loros JJ, Dunlap JC (2019) The phospho-code determining circadian feedback loop closure and output in Neurospora. Mol Cell 74(771–784):e773. https://doi.org/10.1016/j.molcel.2019.03.003
Waterhouse A et al (2018) SWISS-MODEL: homology modelling of protein structures and complexes. Nucleic Acids Res 46:W296–W303. https://doi.org/10.1093/nar/gky427
Wilkins MR, Gasteiger E, Bairoch A, Sanchez JC, Williams KL, Appel RD, Hochstrasser DF (1999) Protein identification and analysis tools in the ExPASy server. Methods Mol Biol 112:531–552. https://doi.org/10.1385/1-59259-584-7:531
Yang J, Zhang Y (2015) I-TASSER server: new development for protein structure and function predictions. Nucleic Acids Res 43:W174-181. https://doi.org/10.1093/nar/gkv342
Yang T, Guo M, Yang H, Guo S, Dong C (2016) The blue-light receptor CmWC-1 mediates fruit body development and secondary metabolism in Cordyceps militaris. Appl Microbiol Biotechnol 100:743–755. https://doi.org/10.1007/s00253-015-7047-6
Funding
This research has been funded by the National Natural Science Foundation of China (Grant No. 31601015) and Foundation of Hebei Educational Committee (Grant No. BJ2016008).
Author information
Authors and Affiliations
Contributions
Qi Xin and Xin Ge performed research, analyzed data, and wrote the paper. Yitong Yuan performed transcriptional analysis. Ruiqing Li prepared and maintained the fungal culture. Xiaomeng Zhang performed PCR and analyzed data. All authors reviewed and corrected the manuscript.
Corresponding author
Ethics declarations
Ethics approval
Not applicable.
Consent to participate
Not applicable.
Consent for publication
Not applicable.
Conflict of interest
The authors declare no competing interests.
Additional information
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Ge, X., Yuan, Y., Li, R. et al. Structure prediction and function characterization of WC-2 proteins in Blakeslea trispora. Int Microbiol 24, 427–439 (2021). https://doi.org/10.1007/s10123-021-00181-1
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10123-021-00181-1