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Plant cholesterol biosynthetic pathway overlaps with phytosterol metabolism

A Corrigendum to this article was published on 12 June 2017


The amount of cholesterol made by many plants is not negligible. Whereas cholesterogenesis in animals was elucidated decades ago, the plant pathway has remained enigmatic. Among other roles, cholesterol is a key precursor for thousands of bioactive plant metabolites, including the well-known Solanum steroidal glycoalkaloids. Integrating tomato transcript and protein co-expression data revealed candidate genes putatively associated with cholesterol biosynthesis. A combination of functional assays including gene silencing, examination of recombinant enzyme activity and yeast mutant complementation suggests the cholesterol pathway comprises 12 enzymes acting in 10 steps. It appears that half of the cholesterogenesis-specific enzymes evolved through gene duplication and divergence from phytosterol biosynthetic enzymes, whereas others act reciprocally in both cholesterol and phytosterol metabolism. Our findings provide a unique example of nature's capacity to exploit existing protein folds and catalytic machineries from primary metabolism to assemble a new, multi-step metabolic pathway. Finally, the engineering of a ‘high-cholesterol’ model plant underscores the future value of our gene toolbox to produce high-value steroidal compounds via synthetic biology.

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Figure 1: The cholesterogenesis pathway in plants and its relationship to phytosterol metabolism and cholesterogenesis in humans.
Figure 2: A transcriptomics and proteomics-based co-expression approach reveals a set of putative cholesterol pathway genes.
Figure 3: Phylogenetic analysis suggests that some of the cholesterol biosynthesis enzymes evolved by duplication and divergence from the phytosterol biosynthesis enzymes.
Figure 4: Distinct pairs of SMOs are involved in cholesterol and phytosterol biosynthesis.
Figure 5: Common enzymes in the cholesterol and phytosterol biosynthetic pathways.
Figure 6: C5-SD2 and 7-DR2 catalyse the last two steps in plant cholesterogenesis.


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We are grateful to D. Twafik for useful suggestions in phylogenetic analysis. A.A. is the incumbent of the Peter J. Cohn Professorial Chair. We thank the Adelis Foundation, the Leona M. and Harry B. Helmsley Charitable Trust, the Jeanne and Joseph Nissim Foundation for Life Sciences, Tom and Sondra Rykoff Family Foundation Research and the Raymond Burton Plant Genome Research Fund for supporting the laboratory activity of A.A. The work was supported by the Israel Science Foundation (ISF Grant No. 1805/15) and the European Research Council (ERC; SAMIT-FP7) personal grants to A.A. P.D.S. is grateful to the Planning and Budgeting Committee of the Council for Higher Education, Israel for the VATAT fellowship. The research in the laboratory of A.G. was financially supported by the VIB International PhD Fellowship Program (fellowship to P.A.) and the Research Foundation Flanders (postdoctoral fellowships to J.P. and L.P.). A.K was supported by a short-term EMBO fellowship (EMBO-ASTF-146-2014). The research in the laboratories of A.A. and A.G. was supported by the European Union Seventh Framework Program FP7/2007–2013 under grant agreement no. 613692–TriForC.

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Authors and Affiliations



P.D.S. designed experiments, performed the research and wrote the paper. J.P., P.A., L.P. and A.G. designed part of the experiments and performed all yeast complementation assays and wrote the paper. S.P. assisted in the VIGS experiments. J.S. and E.S. assisted in the co-expression data analysis. H.M. performed the confocal imaging experiments for localization studies. I.R. and S. Meir assisted with metabolomics data analysis and operated the LCMS. S. Malitsky assisted with GC-S metabolomics data analysis and operated the GCMS. M.Y. and T.U. performed recombinant protein expression in insect cells and isolated microsomes fractions. P.D.C. assisted in wild tomato accessions RNA sequencing. A.M. assisted in sterol extractions and tissue culture work. A.K. and A.P.G. designed part of the research and wrote the paper. H.S. assisted in data analysis and manuscript preparation. A.A. designed the research and wrote the paper.

Corresponding author

Correspondence to Asaph Aharoni.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Information. (PDF 32015 kb)

Supplementary Data 1

Detailed list of genes. (XLSX 18 kb)

Supplementary Data 2

Amino acid sequences. (PDF 286 kb)

Supplementary File 1

Chemical structure: 4α-methyl-5α-cholest-7-en-3βol. (CDX 3 kb)

Supplementary File 2

Chemical structure: 4α-methyl-ergostatrienol. (CDX 3 kb)

Supplementary File 3

Chemical structure: 4α-methyl-24,25-dihydrozymosterol. (CDX 3 kb)

Supplementary File 4

Chemical structure: 4α-methylcholesta-8,14-dien-3β-ol. (CDX 3 kb)

Supplementary File 5

Chemical structure: 4α-methyl-ergostadienol. (CDX 3 kb)

Supplementary File 6

Chemical structure: 7-dehydrocholesterol. (CDX 3 kb)

Supplementary File 7

Chemical structure: 24-ethylidenelophenol. (CDX 3 kb)

Supplementary File 8

Chemical structure: 24-methylenecholesterol. (CDX 3 kb)

Supplementary File 9

Chemical structure: 24-methylenecycloartanol. (CDX 3 kb)

Supplementary File 10

Chemical structure: 24-methylenelophenol. (CDX 3 kb)

Supplementary File 11

Chemical structure: 31-nor-24,25-dihydrolanosterol. (CDX 3 kb)

Supplementary File 12

Chemical structure: 31-norcycloartano. (CDX 3 kb)

Supplementary File 13

Chemical structure: α-chaconine. (CDX 3 kb)

Supplementary File 14

Chemical structure: α-solanine. (CDX 4 kb)

Supplementary File 15

Chemical structure: α-tomatine. (CDX 5 kb)

Supplementary File 16

Chemical structure: β-sitosterol. (CDX 3 kb)

Supplementary File 17

Chemical structure: β-amyrin. (CDX 3 kb)

Supplementary File 18

Chemical structure: campesterol. (CDX 3 kb)

Supplementary File 19

Chemical structure: cholesta-7-en-3β-ol. (CDX 3 kb)

Supplementary File 20

Chemical structure: cholesterol. (CDX 3 kb)

Supplementary File 21

Chemical structure: cycloartanol. (CDX 3 kb)

Supplementary File 22

Chemical structure: cycloartenol. (CDX 3 kb)

Supplementary File 23

Chemical structure: cycloeucalenol. (CDX 3 kb)

Supplementary File 24

Chemical structure ( Delta 5,7 avenasterol) (CDX 3 kb)

Supplementary File 25

chemical structure(Delta 5, Episterol) (CDX 3 kb)

Supplementary File 26

Chemical structure (structure 2): 4,4-dimethylcholesta-8,24-dien-3β-ol (CDX 3 kb)

Supplementary File 27

Chemical structure: δ-7-avenasterol. (CDX 3 kb)

Supplementary File 28

Chemical structure: episterol. (CDX 3 kb)

Supplementary File 29

Chemical structure: esculeoside A. (CDX 7 kb)

Supplementary File 30

Chemical structure: isofucosterol. (CDX 3 kb)

Supplementary File 31

Chemical structure: lanosterol. (CDX 3 kb)

Supplementary File 32

Chemical structure: obtusifoliol. (CDX 3 kb)

Supplementary File 33

Chemical structure: stigmasterol. (CDX 3 kb)

Supplementary File 34

Chemical structure (structure 1): 4,4-dimethylcholesta-8,14(15),24-trien-3β-ol. (CDX 3 kb)

Supplementary File 35

Chemical structure (structure 3): cholesta-8,24-dien-3β-ol. (CDX 3 kb)

Supplementary File 36

Chemical structure (structure 4): cholesta-7,24-dien-3β-ol. (CDX 3 kb)

Supplementary File 37

Chemical structure (structure 5): 7-dehydrodesmosterol. (CDX 3 kb)

Supplementary File 38

Chemical structure (structure 6): desmosterol. (CDX 3 kb)

Supplementary File 39

Chemical structure: uttroside B. (CDX 7 kb)

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Sonawane, P., Pollier, J., Panda, S. et al. Plant cholesterol biosynthetic pathway overlaps with phytosterol metabolism. Nature Plants 3, 16205 (2017).

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