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. 2024 Mar 8;10(10):eadk1992.
doi: 10.1126/sciadv.adk1992. Epub 2024 Mar 8.

A hybrid pathway for self-sustained luminescence

Kseniia A Palkina  1   2 , Tatiana A Karataeva  1   2 , Maxim M Perfilov  1   2 , Liliia I Fakhranurova  1   2 , Nadezhda M Markina  1   2 , Louisa Gonzalez Somermeyer  3 , Elena Garcia-Perez  4 , Marta Vazquez-Vilar  4 , Marta Rodriguez-Rodriguez  4 , Victor Vazquez-Vilriales  4 , Ekaterina S Shakhova  1   2 , Tatiana Mitiouchkina  1   2 , Olga A Belozerova  2 , Sergey I Kovalchuk  2 , Anna Alekberova  1   2 , Alena K Malyshevskaia  1   2 , Evgenia N Bugaeva  1 , Elena B Guglya  2   5 , Anastasia Balakireva  1   2 , Nikita Sytov  2 , Anastasia Bezlikhotnova  1 , Daria I Boldyreva  6 , Vladislav V Babenko  6 , Fyodor A Kondrashov  7 , Vladimir V Choob  8 , Diego Orzaez  4 , Ilia V Yampolsky  1   2   5   9 , Alexander S Mishin  1   2 , Karen S Sarkisyan  1   2   9   10   11
Affiliations

A hybrid pathway for self-sustained luminescence

Kseniia A Palkina et al. Sci Adv. .

Abstract

The fungal bioluminescence pathway can be reconstituted in other organisms allowing luminescence imaging without exogenously supplied substrate. The pathway starts from hispidin biosynthesis-a step catalyzed by a large fungal polyketide synthase that requires a posttranslational modification for activity. Here, we report identification of alternative compact hispidin synthases encoded by a phylogenetically diverse group of plants. A hybrid bioluminescence pathway that combines plant and fungal genes is more compact, not dependent on availability of machinery for posttranslational modifications, and confers autonomous bioluminescence in yeast, mammalian, and plant hosts. The compact size of plant hispidin synthases enables additional modes of delivery of autoluminescence, such as delivery with viral vectors.

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Figures

Fig. 1.
Fig. 1.. Fungal bioluminescence system with plant polyketide synthases.
(A) Caffeic acid cycle catalyzed by nnLuz, nnH3H, nnCPH, and nnHispS (with NpgA) from N. nambi or plant PKS (with 4CL). (B) Protein structures of nnHispS and a plant polyketide synthase (PpASCL), predicted by AlphaFold 2.0 (52). (C) Comparison of luminescence conferred by expression of plant PKS genes in various hosts and relative hispidin concentration in yeast. White space indicates enzymes not included in the corresponding experiment. AMP, adenosine 5′-monophosphate; ATP, adenosine 5′-triphosphate; NAD(P)+, nicotinamide adenine dinucleotide (phosphate); NAD(P)H, reduced form of NAD(P)+; ACP, acyl carrier protein; a.u., arbitrary units.
Fig. 2.
Fig. 2.. Transgenic N. benthamiana plants expressing different versions of the bioluminescence pathway.
(A) Average brightness of leaves, stems, and flowers in 9- to 10-week-old plants. (B) Photo of transgenic plants producing nnHispS, nnLuz, nnH3H, and nnCPH (line NB021). (C and D) Photo of transgenic lines NB221 (C) and NB220 (D) producing PpASCL, nnLuz, nnH3H, and nnCPH. (E) Photo of transgenic plants producing nnHispS, NpgA, nnLuz, nnH3H, and nnCPH (line NB2359). RLU, Relative Luminescence Units.
See this image and copyright information in PMC

References

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