Multiple Cellulose Synthase Catalytic Subunits Are Required for Cellulose Synthesis in Arabidopsis

Taylor, Neil G.; Laurie, Steven S.; Turner, Simon R.

doi:10.2307/3871246

Cited by 95 publications

(164 citation statements)

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“…irx1 and irx3 affect exactly the same cell types and are caused by mutations in the AtCes8 and AtCesA7 genes, respectively. Furthermore, coprecipitation experiments with an epitope-tagged version of IRX3 demonstrate that IRX3 and IRX1 function within the same enzyme complex (9,10). These data are consistent with the idea that at least two CesA gene products are required for cellulose synthesis in both the primary and secondary cell wall of higher plants.…”

supporting

confidence: 82%

“…The region encoding the first variable region of IRX5 (amino acids 96-175) was amplified by PCR with primers IRX5VR1For (5Ј-CGCATATGAATATCAAATATCGCCAGGA-3Ј) and IRX5VR1Rev (5Ј-GCCTCGAGAGTCACTAAACCTCTCT-TCTCT-3Ј) and cloned into the NdeI and XhoI sites of pET24a (Novagen). IRX5VR1 was overexpressed, purified, and used to immunize sheep as described (10), and the antiserum was affinity purified as described.…”

Section: Methodsmentioning

confidence: 99%

“…After transfer to Immuno-blot poly(vinylidine difluoride) membrane (Bio-Rad), protein gel blots were performed according to standard protocols (16). IRX5 and IRX1 antisera were used at 1 in 1,000 dilutions, and IRX3 antisera at 1 in 5,000 (10). Alkalinephosphatase conjugated donkey anti-sheep antibody (Sigma) was used, followed by colorimetric detection of alkaline phosphatase activity to detect antibody binding.…”

Section: Methodsmentioning

confidence: 99%

“…The IRX5 antibody recognizes a band in wild-type extracts corresponding to the correct size for IRX5, indicated by an arrowhead, in addition to bands showing a higher mobility, which are presumed to be degradation products. Rapid degradation of other CesA proteins has been described (10). No band corresponding to IRX5 is seen in irx5-1, which has a 5-kb Ds insertion in IRX5 and presumed to be a null allele, or in irx5-3, which contains a stop codon after 263 aa.…”

Section: Irx5 Is Expressed In the Same Cells As Irx1 And Irx3mentioning

confidence: 99%

“…IRX1 and IRX3 have been shown to associate in the same protein complex, and a hexahistidine tagged IRX3 has been described (10) that may be used for the purification of IRX3 and interacting proteins by using immobilized metal affinity chromatography. IRX5 was solubilized in 2% Triton X-100 (data not shown), the same conditions used to solubilize IRX1 and IRX3 (10). Solubilized stem extract from the hexa-His tagged IRX3 line was bound to nickel resin to purify the tagged IRX3.…”

Section: Irx5 Is Expressed In the Same Cells As Irx1 And Irx3mentioning

confidence: 99%

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Interactions among three distinct CesA proteins essential for cellulose synthesis

Taylor

Howells

Huttly

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

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665

623

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In a screen to identify novel cellulose deficient mutants, three lines were shown to be allelic and define a novel complementation group, irregular xylem5 (irx5). IRX5 was cloned and encodes a member of the CesA family of cellulose synthase catalytic subunits (AtCesA4). irx5 plants have an identical phenotype to previously described mutations in two other members of this gene family (IRX1 and IRX3). IRX5, IRX3, and IRX1 are coexpressed in exactly the same cells, and all three proteins interact in detergent solubilized extracts, suggesting that three members of this gene family are required for cellulose synthesis in secondary cell walls. The association of IRX1 and IRX3 was reduced to undetectable levels in the absence of IRX5. Consequently, these data suggest that IRX5, IRX3, and IRX1 are all essential components of the cellulose synthesizing complex and the presence of all three subunits is required for the correct assembly of this complex.

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supporting

confidence: 82%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Irx5 Is Expressed In the Same Cells As Irx1 And Irx3mentioning

confidence: 99%

Section: Irx5 Is Expressed In the Same Cells As Irx1 And Irx3mentioning

confidence: 99%

See 3 more Smart Citations

Interactions among three distinct CesA proteins essential for cellulose synthesis

Taylor

Howells

Huttly

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

665

623

View full text Add to dashboard Cite

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Genetic dissection of cell wall defects and the strigolactone pathway in Arabidopsis

Ramírez

Pauly

2019

Plant Direct

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Defects in the biosynthesis and/or deposition of secondary plant cell wall polymers result in the collapse of xylem vessels causing a dwarfed plant stature and an altered plant architecture termed irregular xylem ( irx ) syndrome. For example, reduced xylan O ‐acetylation causes strong developmental defects and increased freezing tolerance. Recently, we demonstrated that the irx syndrome in the trichome birefringence‐like 29/eskimo1 ( tbl29/esk1 ) mutant is dependent on MORE AXILLARY GROWTH 4 ( MAX 4), a key enzyme in the biosynthesis of the phytohormone strigolactone ( SL ). In this report, we show that other xylan‐ and cellulose‐deficient secondary wall mutants exhibit increased freezing tolerance correlated with the irx syndrome. In addition, these phenotypes are also dependent on MAX 4, suggesting a more general interaction between secondary wall defects and SL biosynthesis. In contrast, MAX 4 does not play a role in developmental defects triggered by primary wall deficiencies, suggesting that the interaction is restricted to vascular tissue. Through a reverse genetics approach, the requirement of different components of the SL pathway impacting the irx syndrome in tbl29 was evaluated. Our results show that the tbl29 ‐associated irx phenotypes are dependent on the MAX 3 and MAX 4 enzymes, involved in the early steps of SL biosynthesis. In contrast, this signaling is independent on downstream enzymes in the biosynthesis and perception of SL such as MAX 1 and MAX 2.

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Cell wall composition and transcriptomics in stem tissues of stinging nettle (Urtica dioica L.): Spotlight on a neglected fibre crop

et al. 2019

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Stinging nettle ( Urtica dioica L.) produces silky cellulosic fibres, as well as bioactive molecules. To improve the knowledge on nettle and enhance its opportunities of exploitation, a draft transcriptome of the “clone 13” (a fibre clone) is here presented. The transcriptome of whole internodes sampled at the top and middle of the stem is then compared with the core and cortical tissues sampled at the bottom. Young internodes show an enrichment in genes involved in the biosynthesis of phytohormones (auxins and jasmonic acid) and secondary metabolites (flavonoids). The core of internodes collected at the bottom of the stem is enriched in genes partaking in different aspects of secondary cell wall formation (cellulose, hemicellulose, lignin biosynthesis), while the cortical tissues reveal the presence of a C starvation signal probably due to the UDP‐glucose demand necessary for the thickening phase of bast fibres. Cell wall analysis indicates a difference in rhamnogalacturonan structure/composition of mature bast fibres, as evidenced by the higher levels of galactose measured, as well as the occurrence of more water‐soluble pectins in elongating internodes. The targeted quantification of phenolics shows that the middle internode and the cortical tissues at the bottom have higher contents than top internodes. Ultrastructural analyses reveal the presence of a gelatinous layer in bast fibres with a lamellar structure. The data presented will be an important resource and reference for future molecular studies on a neglected fibre crop.

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Multiple Cellulose Synthase Catalytic Subunits Are Required for Cellulose Synthesis in Arabidopsis

Cited by 95 publications

References 0 publications

Interactions among three distinct CesA proteins essential for cellulose synthesis

Interactions among three distinct CesA proteins essential for cellulose synthesis

Genetic dissection of cell wall defects and the strigolactone pathway in Arabidopsis

Cell wall composition and transcriptomics in stem tissues of stinging nettle (Urtica dioica L.): Spotlight on a neglected fibre crop

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