Title: Molecular Studies of the Protein Complexes Involving Cis-Prenyltransferase in Guayule (Parthenium argentatum), an Alternative Rubber-Producing Plant

Abstract

Guayule (Parthenium argentatum) is a perennial shrub in the Asteraceae family and synthesizes a high quality, hypoallergenic cis-1,4-polyisoprene (or natural rubber; NR). Despite its potential to be an alternative NR supplier, the enzymes for cis-polyisoprene biosynthesis have not been comprehensively studied in guayule. Recently, implications of the protein complex involving cis-prenyltransferases (CPTs) and CPT-Binding Proteins (CBPs) in NR biosynthesis were shown in lettuce and dandelion, but such protein complexes have yet to be examined in guayule. In this work, we identified four guayule genes - three PaCPTs (PaCPT1-3) and one PaCBP, whose protein products organize PaCPT/PaCBP complexes. Co-expression of both PaCBP and each of the PaCPTs could complemented the dolichol (a short cis-polyisoprene)-deficient yeast, whereas the individual expressions could not. Microsomes from the PaCPT/PaCBP-expressing yeast efficiently incorporated ¹⁴C-isopentenyl diphosphate into dehydrodolichyl diphosphates; however, NR with high molecular weight could not be synthesized in in vitro assays. Furthermore, co-immunoprecipitation and split-ubiquitin yeast 2-hybrid assays using PaCPTs and PaCBP confirmed the formation of protein complexes. Of the three PaCPTs, guayule transcriptomics analysis indicated that the PaCPT3 is predominantly expressed in stem and induced by cold-stress, suggesting its involvement in NR biosynthesis. The comprehensive analyses of these PaCPTs and PaCBP here provide themore » foundational knowledge to generate a high NR-yielding guayule.« less

Authors:

Lakusta, Adam M. ^[1]; Kwon, Moonhyuk ^[2]; Kwon, Eun-Joo G. ^[1]; Stonebloom, Solomon ^[3]; Scheller, Henrik V. ^[3]; Ro, Dae-Kyun ^[1]

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+ Show Author Affiliations

1. Univ. of Calgary, AB (Canada)
2. Univ. of Calgary, AB (Canada); Gyeongsang National Univ., Jinju (South Korea)
3. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)

Publication Date:

Mon Feb 25 00:00:00 EST 2019

Research Org.:

Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Org.:

USDOE Office of Science (SC), Biological and Environmental Research (BER); National Science and Engineering Research Council of Canada (NSERC); Canada Research Chair (CRC); National Research Foundation of Korea (NRF); e Next-Generation BioGreen 21 Program

OSTI Identifier:

1542354

Grant/Contract Number:  

AC02-05CH11231

Resource Type:

Accepted Manuscript

Journal Name:

Frontiers in Plant Science

Additional Journal Information:

Journal Volume: 10; Journal ID: ISSN 1664-462X

Publisher:

Frontiers Research Foundation

Country of Publication:

United States

Language:

English

Subject:

59 BASIC BIOLOGICAL SCIENCES; terpenoid; cis-prenyltransferase; guayule (Parthenium argentatum); polyisoprene; protein complex