Identification and reconstitution of the rubber biosynthetic machinery on rubber particles from Hevea brasiliensis

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Data availability
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Abstract

Natural rubber (NR) is stored in latex as rubber particles (RPs), rubber molecules surrounded by a lipid monolayer. Rubber transferase (RTase), the enzyme responsible for NR biosynthesis, is believed to be a member of the cis-prenyltransferase (cPT) family. However, none of the recombinant cPTs have shown RTase activity independently. We show that HRT1, a cPT from Hevea brasiliensis, exhibits distinct RTase activity in vitro only when it is introduced on detergent-washed Hevea RPs (WRPs) by a cell-free translation-coupled system. Using this system, a heterologous cPT from Lactuca sativa also exhibited RTase activity, indicating proper introduction of cPT on RP is the key to reconstitute active RTase. RP proteomics and interaction network analyses revealed the formation of the protein complex consisting of HRT1, RUBBER ELONGATION FACTOR (REF) and HRT1-REF BRIDGING PROTEIN. The RTase activity enhancement observed for the complex assembled on WRPs indicates the HRT1-containing complex functions as the NR biosynthetic machinery.

Data availability

The following data sets were generated

1. Takahashi S
2. Nakayama T
(2015) Hevea brasiliensis HRTBP mRNA for transferase binding protein,
Publicly available at the DNA Data Bank of Japan (Accession no: LC057267).

http://www.ddbj.nig.ac.jp/index-e.html

The following previously published data sets were used

1. Qu Y
2. Chakrabarty R
3. Tran HT
4. Kwon EJ
5. Kwon M
6. Nguyen TD
7. Ro DK
(2015) Lactuca sativa cis-prenyltransferase 3 mRNA, complete cds.
Publicly available at the NCBI Nucleotide (Accession no: F752488).

http://www.ncbi.nlm.nih.gov/nuccore/KF752488
1. Rahman AY
2. Usharraj AO
3. Misra BB
4. Thottathil GP
5. Jayasekaran K
6. Feng Y
7. Hou S
8. Ong SY
9. Ng FL
10. Lee LS
11. Tan HS
12. Sakaff MK
13. Teh BS
14. Khoo BF
15. Badai SS
16. Aziz NA
17. Yuryev A
18. Knudsen B
19. Dionne-Laporte A
20. McHunu NP
21. Yu Q
22. Langston BJ
23. Freitas TA
24. Young AG
25. Chen R
26. Wang L
27. Najimudin N
28. Saito JA
29. Alam M
(2013) TSA: Hevea brasiliensis contigxxxxx, mRNA sequence.
Publicly available at the NCBI BioProject (Accession no: PRJNA828950).

https://www.ncbi.nlm.nih.gov/nuccore/?term=PRJNA82895
1. Qu Y
2. Chakrabarty R
3. Tran HT
4. Kwon EJ
5. Kwon M
6. Nguyen TD
7. Ro DK
(2016) Lactuca sativa cis-prenyltransferase-like 2 mRNA, complete cds
Publicly available at the NCBI Nucleotide (Accession no: KF752485).

http://www.ncbi.nlm.nih.gov/nuccore/687843842/

Article and author information

Author details

Satoshi Yamashita

Graduate School of Engineering, Tohoku University, Sendai, Japan

Competing interests
The authors declare that no competing interests exist.
Haruhiko Yamaguchi

Sumitomo Rubber Industries, Ltd, Kobe, Japan

Competing interests
The authors declare that no competing interests exist.
Toshiyuki Waki

Graduate School of Engineering, Tohoku University, Sendai, Japan

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The authors declare that no competing interests exist.
Yuichi Aoki

Graduate School of Information Sciences, Tohoku University, Sendai, Japan

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The authors declare that no competing interests exist.
Makie Mizuno

Graduate School of Engineering, Tohoku University, Sendai, Japan

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The authors declare that no competing interests exist.
Fumihiro Yanbe

Graduate School of Engineering, Tohoku University, Sendai, Japan

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The authors declare that no competing interests exist.
Tomoki Ishii

Graduate School of Engineering, Tohoku University, Sendai, Japan

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The authors declare that no competing interests exist.
Ayuta Funaki

Graduate School of Engineering, Tohoku University, Sendai, Japan

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The authors declare that no competing interests exist.
Yuzuru Tozawa

Graduate School of Science and Engineering, Saitama University, Saitama, Japan

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The authors declare that no competing interests exist.
Yukino Miyagi-Inoue

Sumitomo Rubber Industries, Ltd, Kobe, Japan

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The authors declare that no competing interests exist.
Kazuhisa Fushihara

Sumitomo Rubber Industries, Ltd, Kobe, Japan

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The authors declare that no competing interests exist.
Toru Nakayama

Graduate School of Engineering, Tohoku University, Sendai, Japan

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The authors declare that no competing interests exist.
Seiji Takahashi

Graduate School of Engineering, Tohoku University, Sendai, Japan

For correspondence
takahasi@seika.che.tohoku.ac.jp

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The authors declare that no competing interests exist.

"This ORCID iD identifies the author of this article:" 0000-0002-2288-4340

Funding

No external funding was received for this work.

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This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.