DOI: 10.1126/science.1064281. Sitosterol-β-glucoside as Primer for Cellulose Synthesis in Plants

Sitosterol-β-glucoside as Primer for Cellulose Synthesis in Plants

10.1126/science.1064281

Crossref journal-article

American Association for the Advancement of Science (AAAS)

Science (221)

Abstract

Cellulose synthesis in plants requires β-1,4-glucan chain initiation, elongation, and termination. The process of chain elongation is likely to be distinct from the process of chain initiation. We demonstrate that a CesA glucosyltransferase initiates glucan polymerization by using sitosterol-β-glucoside (SG) as primer. Cotton fiber membranes synthesize sitosterol-cellodextrins (SCDs) from SG and uridine 5′-diphosphate–glucose (UDP-Glc) under conditions that also favor cellulose synthesis. The cellulase encoded by the Korrigan ( Kor ) gene, required for cellulose synthesis in plants, may function to cleave SG from the growing polymer chain.

Bibliography

Peng, L., Kawagoe, Y., Hogan, P., & Delmer, D. (2002). Sitosterol-Î²-glucoside as Primer for Cellulose Synthesis in Plants. Science, 295(5552), 147â150.

Authors 4

Liangcai Peng (first)
Yasushi Kawagoe (additional)
Pat Hogan (additional)
Deborah Delmer (additional)

References 30 Referenced 342

10.1146/annurev.arplant.50.1.245
10.1105/tpc.12.12.2409
10.1104/pp.124.2.495
10.1104/pp.126.1.278
10.1093/pcp/pce045
R. P. Sabba
10.1017/S004317450009144X
10.1073/pnas.191361598
10.1104/pp.126.3.981
L. Peng D. P. Delmer unpublished observations.
10.1074/jbc.274.37.26557
10.1021/bi982531u
10.1074/jbc.271.32.19159
T. Callaghan P. Ross P. Ohana M. Benziman Plant Physiol. 86 1104 (1988). (10.1104/pp.86.4.1104)
10.1046/j.1365-313X.1995.08020213.x
J. L. Cantatore S. M. Murphy D. V. Lynch Biochem. Soc. Trans. 28 part 6 748 (2000). (10.1042/bst0280748)
A. D. Elbein W. T. Forsee Lipids 10 427 (1975). (10.1007/BF02532449)
10.1016/0005-2760(79)90088-2
Fibers (24 days after anthesis) from Gossypium hirsutum Coker 130 were ground in liquid N 2 and extracted at 4°C with 50 mM Na-Mops buffer (or 50 mM Tris-HCl as indicated) pH 7.5 5 mM EDTA 0.25 M sucrose and protease inhibitor cocktail (Boehringer). Extracts were sequentially centrifuged at 2800 g for 20 min then at 100 000 g for 1 hour and the 100 000 g membrane pellet was suspended in 50 mM Mops buffer pH 7.5 containing protease inhibitors. Reaction mixtures containing 5 μM UDP-[ 14 C]Glc (0.5 μCi) 10 mM MgCl 2 2 mM CaCl 2 (or 5 mM EGTA as indicated) 10 mM cellobiose and 250 μg membrane protein in final volume of 0.25 ml were incubated for 1 hour at 37°C and terminated by heating at 100°C for 15 min. Chloroform:methanol (C:M 2:1 0.25 ml) was added with mixing and the resulting upper phase was re-extracted five more times with C:M (2:1). C:M layers were pooled chromatographed on Silica Gel G plates by using C:M:H 2 O (65:25:4) as solvent and subjected to radioautography. [ 14 C]Glucan products in the remaining interface and upper layer were either precipitated at 4°C in 70% ethanol and linkage patterns determined by gas-liquid chromatography (GLC) of permethylated products or by specific enzyme digestions (8).
SG was eluted from TLC plates in chloroform:methanol (C:M 2:1) dried under N 2 and hydrolyzed with 2 M trifluoroacetic acid at 120°C for 1 hour and extracted with C:M:H 2 O (2:1:1). The H 2 O/M phase was collected for sugar analysis by GLC (27) and the C:M phase was used for identification of the major sterol as sitosterol by retention time and mass spectrum with GLC/MS (DB-17 capillary column). A molecular ion ( m/z 414) and prominent fragment ions diagnostic of sitosterol ( m/z 255 281 303 329 381 396) were observed. We also determined by GLC that the fatty acid acylated to SG to form ASG is palmitic acid .
Supplementary Web material is available on Science Online at www.sciencemag.org/cgi/content/full/295/5552/147/DC1.
10.1104/pp.010269
10.1073/pnas.94.17.9091
Schrick K., et al., Genes Dev. 14, 1471 (2000). (10.1101/gad.14.12.1471) / Genes Dev. by Schrick K. (2000)
10.1046/j.1365-313x.2001.00994.x
N. G. Taylor S. Laurie S. R. Turner Plant Cell 12 2529 (2000). (10.2307/3871246)
10.1016/S0960-9822(01)00108-7
10.1016/0008-6215(83)88244-5
10.1016/S0003-2697(69)80009-6
This work was supported by grant DE-FG-03-963ER 20238 from the U.S. Department of Energy. We thank K. Kreuz of Syngenta Basel Switzerland for the gift of CGA 325'615 and A. Bennett of University of California at Davis for antibody against tomato Cel3 (Kor).

Dates

Type	When
Created	23 years, 1 month ago (July 27, 2002, 5:47 a.m.)
Deposited	1 year, 7 months ago (Jan. 9, 2024, 6:08 p.m.)
Indexed	5 days, 5 hours ago (Aug. 24, 2025, 6:55 p.m.)
Issued	23 years, 7 months ago (Jan. 4, 2002)
Published	23 years, 7 months ago (Jan. 4, 2002)
Published Print	23 years, 7 months ago (Jan. 4, 2002)

Funders 0

None

BibTeX

@article{Peng_2002, title={Sitosterol-β-glucoside as Primer for Cellulose Synthesis in Plants}, volume={295}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.1064281}, DOI={10.1126/science.1064281}, number={5552}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Peng, Liangcai and Kawagoe, Yasushi and Hogan, Pat and Delmer, Deborah}, year={2002}, month=jan, pages={147–150} }

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  "abstract": "<jats:p>\n            Cellulose synthesis in plants requires \u03b2-1,4-glucan chain initiation, elongation, and termination. The process of chain elongation is likely to be distinct from the process of chain initiation. We demonstrate that a CesA glucosyltransferase initiates glucan polymerization by using sitosterol-\u03b2-glucoside (SG) as primer. Cotton fiber membranes synthesize sitosterol-cellodextrins (SCDs) from SG and uridine 5\u2032-diphosphate\u2013glucose (UDP-Glc) under conditions that also favor cellulose synthesis. The cellulase encoded by the\n            <jats:italic>Korrigan</jats:italic>\n            (\n            <jats:italic>Kor</jats:italic>\n            ) gene, required for cellulose synthesis in plants, may function to cleave SG from the growing polymer chain.\n          </jats:p>",
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