Genome-wide identification and analysis of AP2/ERF transcription factors related to camptothecin biosynthesis in Camptotheca acuminata

doi:10.1016/S1875-5364(20)30070-4

Chinese Journal of Natural Medicines

Volume 18, Issue 8, August 2020, Pages 582-593

https://doi.org/10.1016/S1875-5364(20)30070-4 Get rights and content

Abstract

Camptotheca acuminata produces camptothecin (CPT), a monoterpene indole alkaloid (MIA) that is widely used in the treatment of lung, colorectal, cervical, and ovarian cancers. Its biosynthesis pathway has attracted significant attention, but the regulation of CPT biosynthesis by the APETALA2/ethylene-responsive factor (AP2/ERF) transcription factors (TFs) remains unclear. In this study, a systematic analysis of the AP2/ERF TFs family in C. acuminata was performed, including phylogeny, gene structure, conserved motifs, and gene expression profiles in different tissues and organs (immature bark, cotyledons, young flower, immature fruit, mature fruit, mature leaf, roots, upper stem, and lower stem) of C. acuminata. A total of 198 AP2/ERF genes were identified and divided into five relatively conserved subfamilies, including AP2 (26 genes), DREB (61 genes), ERF (92 genes), RAV (18 genes), and Soloist (one gene). The combination of gene expression patterns in different C. acuminata tissues and organs, the phylogenetic tree, the co-expression analysis with biosynthetic genes, and the analysis of promoter sequences of key enzymes genes involved in CPT biosynthesis pathways revealed that eight AP2/ERF TFs in C. acuminata might be involved in CPT synthesis regulation, which exhibit relatively high expression levels in the upper stem or immature bark. Among these, four genes (CacAP2/ERF123, CacAP2/ERF125, CacAP2/ERF126, and CacAP2/ERF127) belong to the ERF–B2 subgroup; two genes (CacAP2/ERF149 and CacAP2/ERF152) belong to the ERF–B3 subgroup; and two more genes (CacAP2/ERF095 and CacAP2/ERF096) belong to the DREB–A6 subgroup. These results provide a foundation for future functional characterization of the AP2/ERF genes to enhance the biosynthesis of CPT compounds of C. acuminata.

References (56)

M Pfaffenbach et al.
The rhazinilam-leuconoxine-mersicarpine triad of monoterpenoid indole alkaloids
Alkaloids Chemi Biol
(2017)
A Lorence et al.
Camptothecin, over four decades of surprising findings
Phytochem
(2004)
M Wink
Evolution of secondary metabolites from an ecological and molecular phylogenetic perspective
Phytochem
(2003)
YH Hsiang et al.
Camptothecin induces protein-linked DNA breaks via mammalian DNA topo I
J Biol Chem
(1985)
S Sirikantaramas et al.
Camptothecin: biosynthesis, biotechnological production and resistance mechanism(s)
Adv Bot Res
(2013)
CY Li et al.
The ORCA2 transcription factor plays a key role in regulation of the terpenoid indole alkaloid pathway
BMC Plant Biol
(2013)
Y Zhang et al.
The AP2/ERF transcription factor SmERF128 positively regulates diterpenoid biosynthesis in Salvia miltiorrhiza
Plant Mol Biol
(2019)
Q Huang et al.
The AP2/ERF transcription factor SmERF1L1 regulates the biosynthesis of tanshinones and phenolic acids in Salvia miltiorrhiza
Food Chem
(2019)
ZX Yu et al.
The jasmonate-responsive AP2/ERF transcription factors AaERF1 and AaERF2 positively regulate artemisinin biosynthesis in Artemisia annua L
Mol Plant
(2012)
SR Wessler
Homing into the origin of the AP2 DNA binding domain
Trends Plant Sci
(2005)

Y Sakuma et al.

DNA-binding specificity of the ERF/AP2 domain of arabidopsis DREBs, transcription factors involved in dehydration- and cold-inducible gene expression

Biochem Biophys Res Commun

(2002)

E Gongora-Castillo et al.

Genomic approaches for interrogating the biochemistry of medicinal plant species

Methods Enzymol

(2012)

LV De et al.

Making iridoids/secoiridoids and monoterpenoid indole alkaloids: progress on pathway elucidation

Curr Opin Plant Biol

(2014)

TM Kutchan

Alkaloid biosynthesis: the basis for metabolic engineering of medicinal plants

Plant Cell

(1995)

RU Shaanker et al.

Chemical Profiling of Nothapodytes nimmoniana for Camptothecin, An Important Anticancer Alkaloid: Towards the Development of a Sustainable Production System [M]. Bioactive Molecules and Medicinal Plants

(2008)

N Udomsom et al.

Function of AP2/ERF transcription factors involved in the regulation of specialized metabolism in Ophiorrhiza pumila revealed by transcriptomics and metabolomics

Front Plant Sci

(2016)

J Garst

Topotecan: an evolving option in the treatment of relapsed small cell lung cancer

Ther Clin Risk Manag

(2008)

D Cunningham et al.

Optimizing the use of irinotecan in colorectal cancer

Oncologist

(2001)

RL Coleman

Emerging role of topotecan in front-line treatment of carcinoma of the ovary

Oncologist

(2002)

C Chang et al.

A bZIP transcription factor, CaLMF, mediated light-regulated camptothecin biosynthesis in Camptotheca acuminata

Tree Physiol

(2019)

Y Yamazaki et al.

Biosynthesis of camptothecin. In silico and in vivotracer study from [1–13C] glucose

Plant Physiol

(2004)

GY Kai et al.

Biosynthesis and biotechnological production of anti-cancer drug camptothecin

Phytochem Rev

(2015)

GA Rather et al.

Biosynthetic system of camptothecin: an anticancer plant product

Pure Appl Chem

(2010)

G Kai et al.

Effect of three plant hormone elicitors on the camptothecin accumulation and gene transcript profiling in Camptotheca acuminata seedlings

Inter J Sci

(2014)

R Sadre et al.

Metabolite diversity in alkaloid biosynthesis: a multilane (diastereomer) highway for camptothecin synthesis in Camptotheca acuminata

Plant Cell

(2016)

Y Yang et al.

Bifunctional cytochrome P450 enzymes involved in camptothecin biosynthesis

ACS Chem Biol

(2019)

V Salim et al.

Camptotheca acuminata 10-hydroxycamptothecin O-methyltransferase: an alkaloid biosynthetic enzyme co-opted from flavonoid metabolism

Plant J

(2018)

RC Elliott et al.

Aintegumenta, an Apetala2-like gene of arabidopsis with pleiotropic roles in ovule development and floral organ growth

Plant Cell

(1996)

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Camptotheca acuminata is one of the primary sources of camptothecin (CPT), which is widely used in the treatment of human malignancies because of its inhibitory activity against DNA topoisomerase I. Although several transcription factors have been identified for regulating CPT biosynthesis in other species, such as Ophiorrhiza pumila, the specific regulatory components controlling CPT biosynthesis in C. acuminata have yet to be definitively determined. In this study, CaERF1, an DREB subfamily of the APETALA2/ethylene response factors (AP2ERFs), was identified in C. acuminata. The transient overexpression and silencing of CaERF1 in C. acuminata leaves confirmed that it positively regulates the accumulation of CPT by inducing the expression of CaCYC1 and CaG8O in the iridoid pathway. Results of transient transcriptional activity assay and yeast one-hybrid assays have showed that CaERF1 transcriptionally activates the expression of CaCYC1 and CaG8O by binding to RAA and CEI elements in the promoter regions of these two genes. Furthermore, the expression of CaCYC1 and CaG8O in CaERF1-silenced leaves was less sensitive to ABA treatment, indicating that CaERF1 is a crucial component involved in ABA-regulated CPT biosynthesis in C. acuminata.
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Functional verification analysis found that overexpression of CaLMF inhibited the camptothecin accumulation, and silencing of CaLMF abolished the regulation of camptothecin by light, indicating that CaLMF is an important factor mediating light-regulated camptothecin biosynthesis (Chang et al., 2019). Comparative analysis of AP2/ERF transcription factors in C. acuminata genome revealed that a total of 198 genes were classified into 5 subfamilies and 8 candidate genes that might be involved in the regulation of camptothecin biosynthesis were identified (Hu et al., 2020). Furthermore, overexpression of the exogenous ORCA3 transcription factor from C. roseus in C. acuminata hairy roots could increase camptothecin content by approximately 1.5-fold (Ni et al., 2011).
Camptothecin (CPT), a well-known natural anti-cancer drug, is produced from woody plant Camptotheca acuminata. Up to now, two CPT-derivatives, irinotecan and topotecan, have been used to treat various cancers all over the world, and more than a dozen camptothecin analogues are in different stages of clinical research and development. The source of camptothecin mainly depends on the extraction from the fruits and barks of C. acuminata to meet the huge clinical demand, which undoubtedly increases the logging of this CPT-producing plant resource. Therefore, it is crucial to improve the content of camptothecin in C. acuminata using metabolic engineering strategies for the sustainable development of medicinal source plants. The camptothecin biosynthesis pathway is complex and only the functional genes that catalyze the biosynthesis of central intermediate strictosidinic acid and CPT-derivatives have been partially resolved. Thus, this review mainly summarized the anti-tumor mechanism and application of camptothecin drugs, the tissue differential distribution and extraction of camptothecin, the biosynthetic genes of camptothecin and its derivatives in C. acuminate. In addition, the application of camptothecin metabolic engineering in CPT-producing plants was also prospected. This review provided assistance for increasing camptothecin content and C. acuminata germplasm innovation using camptothecin metabolic engineering technologies.
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As AP2/ERF superfamily TFs have important roles against diverse stresses, an in-depth study of this TFs may help to get stress tolerance mechanisms of the jute plant. The AP2/ERF superfamily genes have been investigated in diverse plant species, including Arabidopsis (Nakano et al., 2006), rice (Rashid et al., 2012), buckwheat (Liu et al., 2019), pineapple (Huang et al., 2020), sorghum (Mathur et al., 2020), Camptotheca acuminata (Ya-Ting et al., 2020), Andrographis paniculata (Yao et al., 2020), ginseng (Chen et al., 2020), orchardgrass (Xu et al., 2020a,b), and Gynostemma pentaphyllum (Xu et al., 2020a,b). To our best knowledge, no systematic investigation on the AP2/ERF superfamily genes in jute has been accomplished.
The ethylene-responsive element (AP2/ERF) is one of the keys and conserved transcription factors (TF) in plants that play a vital role in regulating plant growth, development and stress response. A total of 119 CoAP2/ERF genes were identified from the dark jute genome, one of the most important bast fibre crops. Among these, 17 were AP2, 97 ERF, 4 RAV and 1 Soloist gene family. Phylogenetic analysis revealed that the AP2/ERF genes can be divided into 10 groups. These genes were distributed randomly across the linkage groups and found three tandem and eight segmental duplications which arose from ancient duplication events. Expression profiling of CoAP2/ERF genes was assessed using RNA-Seq data and qRT-PCR analysis in tissues during development and under biotic (Macrophomina phaseolina) and abiotic (waterlogging, salinity, drought) stresses. CoERF-21, CoERF-34 and CoERF-39 genes were significantly upregulated during waterlogging stress and CoDREB-11, CoDREB-14 and CoRAV-01 genes were upregulated under salinity and drought stress condition. Gene evolution analysis among the Malvaceae family showed dark jute were separated 14 and 22 Mya from cacao and cotton respectively and expression correlation analysis revealed the functions of superfamily genes were expanded and enhanced through gene duplication. These findings provide new insights into AP2/ERF gene evolution, divergence in dark jute and considered a valuable resource for further functional validation as well as for utilization in stress resistance variety development program.

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Available online 20 Aug., 2020

Research funding This work was supported by the National Key R&D Program of China (No. 2019YFC1711100), and the CAMS Innovation Fund for Medical Sciences (CIFMS, No. 2016-I2M-3-016)

These authors have no conflict of interest to declare.

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Special topicGenome-wide identification and analysis of AP2/ERF transcription factors related to camptothecin biosynthesis in Camptotheca acuminata

Abstract

Alkaloids Chemi Biol

Phytochem

Phytochem

J Biol Chem

Adv Bot Res

BMC Plant Biol

Plant Mol Biol

Food Chem

Mol Plant

Trends Plant Sci

Biochem Biophys Res Commun

Methods Enzymol

Making iridoids/secoiridoids and monoterpenoid indole alkaloids: progress on pathway elucidation

Curr Opin Plant Biol

Alkaloid biosynthesis: the basis for metabolic engineering of medicinal plants

Plant Cell

Chemical Profiling of Nothapodytes nimmoniana for Camptothecin, An Important Anticancer Alkaloid: Towards the Development of a Sustainable Production System [M]. Bioactive Molecules and Medicinal Plants

Function of AP2/ERF transcription factors involved in the regulation of specialized metabolism in Ophiorrhiza pumila revealed by transcriptomics and metabolomics

Front Plant Sci

Topotecan: an evolving option in the treatment of relapsed small cell lung cancer

Ther Clin Risk Manag

Optimizing the use of irinotecan in colorectal cancer

Oncologist

Emerging role of topotecan in front-line treatment of carcinoma of the ovary

Oncologist

A bZIP transcription factor, CaLMF, mediated light-regulated camptothecin biosynthesis in Camptotheca acuminata

Tree Physiol

Biosynthesis of camptothecin. In silico and in vivotracer study from [1–13C] glucose

Plant Physiol

Biosynthesis and biotechnological production of anti-cancer drug camptothecin

Phytochem Rev

Biosynthetic system of camptothecin: an anticancer plant product

Pure Appl Chem

Effect of three plant hormone elicitors on the camptothecin accumulation and gene transcript profiling in Camptotheca acuminata seedlings

Inter J Sci

Metabolite diversity in alkaloid biosynthesis: a multilane (diastereomer) highway for camptothecin synthesis in Camptotheca acuminata

Plant Cell

Bifunctional cytochrome P450 enzymes involved in camptothecin biosynthesis

ACS Chem Biol

Camptotheca acuminata 10-hydroxycamptothecin O-methyltransferase: an alkaloid biosynthetic enzyme co-opted from flavonoid metabolism

Plant J

Aintegumenta, an Apetala2-like gene of arabidopsis with pleiotropic roles in ovule development and floral organ growth

Plant Cell

Special topic
Genome-wide identification and analysis of AP2/ERF transcription factors related to camptothecin biosynthesis in Camptotheca acuminata