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Comprehensive characterization of Guanosine monophosphate synthetase in Nicotiana tabacum

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Abstract

Background

Guanosine monophosphate (GMP) synthetase is an enzyme that converts xanthosine monophosphate to GMP. GMP plays an essential role in plant development and responses to internal and external stimuli. It also plays a crucial role in several plant physiochemical processes, such as stomata closure, cation flux regulation, pathogen responses and chloroplast development.

Methods and results

The mRNA sequences of NtGMP synthase in tobacco (Nicotiana tabacum) were rapidly amplified from cDNA. The GMP synthase open reading frame contains a 1617 bp sequence encoding 538 amino acids. A sequence analysis showed that this sequence shares high homology with that of Nicotiana sylvestris, Nicotiana attenuata, N. tomentosiformis, Solanum tuberosum, Lycopersicon pennellii, L. esculentum, Capsicum annuum, C. chinense and C. baccatum GMP synthase. A BLAST analysis with a tobacco high-throughput genomic sequence database revealed that the tobacco GMP synthase gene has five introns and six exons. A phylogenetic analysis showed a close genetic evolutionary relationship with N. sylvestris GMP synthase. The tissue-specific expression profile was evaluated using quantitative real-time PCR. The data showed that NtGMP synthase was highly expressed in leaves and moderately expressed in roots, flowers, and stems. The subcellular localization was predicted using the WOLF PSORT webserver, which strongly suggested that it might be localized to the cytoplasm.

Conclusions

In the current study, we cloned and comprehensively characterized GMP synthase in tobacco (Nicotiana tabacum). Our results establish a basis for further research to explore the precise role of this enzyme in tobacco.

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References

  1. Nadimpalli R, Yalpani N, Johal GS, Simmons CR (2000) Prohibitins, stomatins, and plant disease response genes compose a protein superfamily that controls cell proliferation, ion channel regulation, and death. J Biol Chem 275:29579–29586

    Article  CAS  Google Scholar 

  2. Abbott JL et al (2006) The effects of removing the GAT domain from E. coli GMP synthetase. Protein J 25:483–491

    Article  CAS  Google Scholar 

  3. Li Q et al (2015) GMP synthase is essential for viability and infectivity of T rypanosoma brucei despite a redundant purine salvage pathway. Mol Microbiol 97:1006–1020

    Article  CAS  Google Scholar 

  4. Welin M et al (2013) Substrate specificity and oligomerization of human GMP synthetase. J Mol Biol 425:4323–4333

    Article  CAS  Google Scholar 

  5. Wang Q, Guan YF, Hancock SE, Wahi K, van Geldermalsen M, Zhang BK, Pang A, Nagarajah R, Mak B, Freidman N, Horvath LG (2021) Inhibition of guanosine monophosphate synthetase (GMPS) blocks glutamine metabolism and prostate cancer growth. J Pathol 254:135–146

    Article  CAS  Google Scholar 

  6. Ko TP, Wang YC, Tsai CL, Yang CS, Hou MH, Chen Y (2021) Crystal structure and functional implication of a bacterial cyclic AMP–AMP–GMP synthetase. Nucleic Acids Res 49:4725–4737

    Article  CAS  Google Scholar 

  7. Townsley L, Yannarell SM, Huynh TN, Woodward JJ, Shank EA (2018) Cyclic di-AMP acts as an extracellular signal that impacts Bacillus subtilis biofilm formation and plant attachment. MBio 9:e00341–e00318

    Article  CAS  Google Scholar 

  8. Herz S, Eberhardt S, Bacher A (2000) Biosynthesis of riboflavin in plants. The ribA gene of Arabidopsis thaliana specifies a bifunctional GTP cyclohydrolase II/3, 4-dihydroxy-2-butanone 4-phosphate synthase. Phytochem 53:723–731

    Article  CAS  Google Scholar 

  9. Moffatt BA, Ashihara H (2002) Purine and pyrimidine nucleotide synthesis and metabolism. The Arabidopsis Book/American Society of Plant Biologists 1:e0018

  10. Boldt R, Zrenner R (2003) Purine and pyrimidine biosynthesis in higher plants. Physiol Planta 117:297–304

    Article  CAS  Google Scholar 

  11. Zrenner R, Stitt M, Sonnewald U, Boldt R (2006) Pyrimidine and purine biosynthesis and degradation in plants. Annu Rev Plant Biol 57:805–836

    Article  CAS  Google Scholar 

  12. Hove-Jensen B et al (2017) Phosphoribosyl diphosphate (PRPP): biosynthesis, enzymology, utilization, and metabolic significance. Microbiol Mol Biol Rev 81:e00040-16

    Article  CAS  Google Scholar 

  13. Abbas F, Ke Y, Yu R, Fan Y (2019) Functional characterization and expression analysis of two terpene synthases involved in floral scent formation in Lilium ‘Siberia.’ Planta 249:71–93

    Article  CAS  Google Scholar 

  14. Abbas F et al (2020) Cloning, functional characterization and expression analysis of LoTPS5 from Lilium ‘Siberia’. Gene 756:144921

  15. Ke Y et al (2019) Genome-wide analysis and characterization of the Aux/IAA Family genes related to floral scent formation in Hedychium coronarium. Inter J Mol Sci 20:3235

    Article  CAS  Google Scholar 

  16. Ke Y, Abbas F, Zhou Y, Yu R, Fan Y (2021) Auxin-responsive R2R3-MYB transcription factors HcMYB1 and HcMYB2 activate volatile biosynthesis in Hedychium coronarium flowers. Front Plant Sci 12:1626

    Google Scholar 

  17. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 25:402–408

    Article  CAS  Google Scholar 

  18. Larkin MA et al (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948

    Article  CAS  Google Scholar 

  19. Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874

    Article  CAS  Google Scholar 

  20. Letunic I, Bork P (2019) Interactive Tree Of Life (iTOL) v4: recent updates and new developments. Nucleic Acids Res 47:W256–W259

    Article  CAS  Google Scholar 

  21. Stothard P (2000) The sequence manipulation suite: JavaScript programs for analyzing and formatting protein and DNA sequences. Biotechniques 28:1102–1104

    Article  CAS  Google Scholar 

  22. Lescot M et al (2002) PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Res 30:325–327

    Article  CAS  Google Scholar 

  23. Ballut L, Violot S, Shivakumaraswamy S, Thota LP, Sathya M, Kunala J, Dijkstra BW, Terreux R, Haser R, Balaram H, Aghajari N (2015) Active site coupling in Plasmodium falciparum GMP synthetase is triggered by domain rotation. Nature Commun 6:1–13

    Article  Google Scholar 

  24. Rodriguez-Suarez R et al (2007) Mechanism-of-action determination of GMP synthase inhibitors and target validation in Candida albicans and Aspergillus fumigatus. Chem Biol 14:1163–1175

    Article  CAS  Google Scholar 

  25. Wei-Fon H, Chen LJ, Boldt R, Sun CW, Li HM (2004) Characterization of Arabidopsis Glutamine Phosphoribosyl Pyrophosphate Amidotransferase-Deficient Mutants1. Plant Physiol 135:1314

    Article  Google Scholar 

  26. Ennajdaoui H et al (2010) Trichome specific expression of the tobacco (Nicotiana sylvestris) cembratrien-ol synthase genes is controlled by both activating and repressing cis-regions. Plant Mol Biol 73:673–685

    Article  CAS  Google Scholar 

  27. Katahira R, Ashihara H (2006) Profiles of purine biosynthesis, salvage and degradation in disks of potato (Solanum tuberosum L.) tubers. Planta 225:115–126

    Article  CAS  Google Scholar 

  28. Li-Ping Z, Han-Xia L, Ouyang B, Zhang JH, Zhi-Biao Y (2006) Cloning, Expression, and Mapping of GDP-D-mannose Pyrophosphorylase cDNA from Tomato (Lycopersicon esculentum). Acta Genet Sinica 33:757–764

    Article  Google Scholar 

  29. Gross I, Durner J (2016) In search of enzymes with a role in 3′, 5′-cyclic guanosine monophosphate metabolism in plants. Front Plant Sci 7:576

    Article  Google Scholar 

  30. Xiong ZQ, Fan YZ, Song X, Liu XX, Xia YJ, Ai LZ (2020) The second messenger c-di-AMP mediates bacterial exopolysaccharide biosynthesis: a review. Mol Biol Rep 47:9149–9157

    Article  CAS  Google Scholar 

  31. Maruoka S, Horita S, Lee WC, Nagata K, Tanokura M (2010) Crystal structure of the ATPPase subunit and its substrate-dependent association with the GATase subunit: a novel regulatory mechanism for a two-subunit-type GMP synthetase from Pyrococcus horikoshii OT3. J Bol Biol 395:417–429

    Article  CAS  Google Scholar 

  32. Rahman H, Wang XY, Xu YP, He YH, Cai XZ (2020) Characterization of tomato protein kinases embedding guanylate cyclase catalytic center motif. Sci Rep 10:1–16

    Article  Google Scholar 

  33. Sonawane AR et al (2017) Understanding tissue-specific gene regulation. Cell Rep 21:1077–1088

    Article  CAS  Google Scholar 

  34. He J, Ruan W, Sun J, Wang F, Yan W (2018) Functional characterization of c-di-GMP signaling-related genes in the probiotic Lactobacillus acidophilus. Front Microbiol 9:1935

    Article  Google Scholar 

  35. Liu L et al (2015) A resource for characterizing genome-wide binding and putative target genes of transcription factors expressed during secondary growth and wood formation in Populus. Plant J 82:887–898

    Article  CAS  Google Scholar 

Download references

Funding

This work was supported by the Science and Technology Plan Project of Yunnan Provincial Tobacco Company (2018530000241016, 2018530000241020, 2019530000241028, and 2019530000241011), Shanghai Tobacco Group Co., Ltd. (20193100001 and 20203100001), Yunnan Provincial Department of Science and Technology (2019FH001(-051)) and Science and Technology Project of Yunnan Provincial Education Department (2021J0716).

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Authors and Affiliations

  1. College of Agriculture and Life Sciences, Yunnan Urban Agricultural Engineering & Technological Research Center, Kunming University, Kunming, China

    Feiyan Huang, Ke Yanguo & Lei Yu

  2. The Research Center for Ornamental Plants, College of Forestry and Landscape Architecture, South China Agricultural University, Guangzhou, 510642, China

    Farhat Abbas

  3. Department of Plant Breeding and Genetics, University of Haripur, Khyber Pakhtunkhwa, Pakistan

    Sajid Fiaz

  4. Department of Crop Science and Technology, College of Agriculture, South China Agricultural University, Guangzhou, 510642, China

    Muhammad Imran

  5. College of Economics and Management, Kunming University, Kunming, China

    Ke Yanguo

  6. Institute of Environment and Sustainable Development in Agricultural, Chinese Academy of Agricultural Sciences, Beijing, China

    Waseem Hassan

  7. Department of Botany, Division of Science and Technology, University of Education Lahore, Punjab, Pakistan

    Umair Ashraf

  8. Lincang Tobacco Corporation of Yunnan Province, Kunming, China

    Yuansheng He

  9. Material Procurement Center, Shanghai Tobacco Group Co., Ltd, Shanghai, 200082, China

    Xuanjie Cai

  10. Kunming Tobacco Corporation of Yunnan Province, Kunming, 650021, China

    Zhijiang Wang & Xianwen Ye

  11. Tobacco Leaf Technology Centre, China Tobacco Henan Industrial Co., Ltd, Zhengzhou, 450000, China

    Xiaolong Chen

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  1. Feiyan Huang
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Corresponding authors

Correspondence to Ke Yanguo, Xianwen Ye or Xiaolong Chen.

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Supplementary Information

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Supplementary Figure 1: Prediction of transmembrane helices in the NtGMP synthase protein (gif 4 kb)

11033_2021_6718_MOESM2_ESM.xlsx

Supplementary Table 1: List of NtGMP synthase primers used in the assays. Supplementary Table 2: Accession numbers of GMP synthases retrieved from various plant species. Supplementary Table 3: In silico cis-regulatory elements present in the tobacco GMP synthase sequence (XLSX 21 kb)

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Huang, F., Abbas, F., Fiaz, S. et al. Comprehensive characterization of Guanosine monophosphate synthetase in Nicotiana tabacum. Mol Biol Rep 49, 5265–5272 (2022). https://doi.org/10.1007/s11033-021-06718-x

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  • DOI: https://doi.org/10.1007/s11033-021-06718-x

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