Abstract
Cellulose is a major component of the extracellular matrices formed during development of the social amoeba, Dictyostelium discoideum. We isolated insertional mutants that failed to accumulate cellulose and had no cellulose synthase activity at any stage of development. Development proceeded normally in the null mutants up to the beginning of stalk formation, at which point the culminating structures collapsed onto themselves, then proceeded to attempt culmination again. No spores or stalk cells were ever made in the mutants, with all cells eventually lysing. The predicted product of the disrupted gene (dcsA) showed significant similarity to the catalytic subunit of cellulose synthases found in bacteria. Enzyme activity and normal development were recovered in strains transformed with a construct expressing the intact dcsA gene. Growing amoebae carrying the construct accumulated the protein product of dcsA, but did not make cellulose until they had developed for at least 10 hr. These studies show directly that the product of dcsA is necessary, but not sufficient, for synthesis of cellulose.
MeSH Terms
Amino Acid Sequence
Animals
Base Sequence
Catalytic Domain
Cellulose/biosynthesis
Dictyostelium/enzymology,genetics
Gene Expression
Glucosyltransferases/genetics
Molecular Sequence Data
Mutagenesis
Phenotype
Protozoan Proteins
Chemicals
Protozoan Proteins
Cellulose
DcsA protein, Dictyostelium
Glucosyltransferases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Blanton R L
Department of Biological Sciences, Texas Tech University, Lubbock, TX 79409, USA.
[email protected]
Fuller D
Iranfar N
Grimson M J
Loomis W F
References (28)
28 references, click to expand
-
Simple apparatus for plunge-freezing biological material: Some design considerations.
Microsc Res Tech. 1999 Jan 15-Feb 1;44(2-3):195-200
PMID: 10084826
-
The phosphodiesterase secreted by prestalk cells is necessary for Dictyostelium morphogenesis.
Dev Biol. 1995 Jan;167(1):1-8
PMID: 7851634
-
Cultivation and synchronous morphogenesis of Dictyostelium under controlled experimental conditions.
Methods Cell Biol. 1987;28:9-29
PMID: 3298997
-
Polysaccharides of Chlorella pyrenoidosa.
Biochem J. 1962 Mar;82:509-19
PMID: 14481680
-
In vitro glucan synthesis by membranes of celery petioles: the role of the membrane in determining the type of linkage formed.
J Cell Sci Suppl. 1985;2:1-11
PMID: 2936754
-
Genes required for cellulose synthesis in Agrobacterium tumefaciens.
J Bacteriol. 1995 Feb;177(4):1069-75
PMID: 7860585
-
DNA sequences required for expression of a Dictyostelium actin gene.
EMBO J. 1986 Dec 1;5(12):3361-6
PMID: 3028782
-
New approaches to the study of cellulose biosynthesis.
J Cell Sci Suppl. 1985;2:33-50
PMID: 2419349
-
Use of a fluorescent brightener to demonstrate cellulose in the cellular slime molds.
Appl Microbiol. 1968 Jan;16(1):106-13
PMID: 5688830
-
Activity of the yeast MNN1 alpha-1,3-mannosyltransferase requires a motif conserved in many other families of glycosyltransferases.
Proc Natl Acad Sci U S A. 1998 Jul 7;95(14):7945-50
PMID: 9653120
-
Antisense RNA inactivation of myosin heavy chain gene expression in Dictyostelium discoideum.
Science. 1987 May 29;236(4805):1081-6
PMID: 3576221
-
Multidomain architecture of beta-glycosyl transferases: implications for mechanism of action.
J Bacteriol. 1995 Mar;177(6):1419-24
PMID: 7883697
-
Cloning in silico.
Curr Biol. 1997 Feb 1;7(2):R108-11
PMID: 9081659
-
Isolation of Dictyostelium discoideum cytokinesis mutants by restriction enzyme-mediated integration of the blasticidin S resistance marker.
Biochem Biophys Res Commun. 1994 Dec 30;205(3):1808-14
PMID: 7811269
-
Higher plants contain homologs of the bacterial celA genes encoding the catalytic subunit of cellulose synthase.
Proc Natl Acad Sci U S A. 1996 Oct 29;93(22):12637-42
PMID: 8901635
-
Cloning and sequencing of the cellulose synthase catalytic subunit gene of Acetobacter xylinum.
Plant Mol Biol. 1990 Nov;15(5):673-83
PMID: 2151718
-
Identification of a second cellulose synthase gene (acsAII) in Acetobacter xylinum.
J Bacteriol. 1995 Sep;177(18):5276-83
PMID: 7665515
-
REMI-RFLP mapping in the Dictyostelium genome.
Genetics. 1994 Nov;138(3):665-74
PMID: 7851764
-
Tagging developmental genes in Dictyostelium by restriction enzyme-mediated integration of plasmid DNA.
Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8803-7
PMID: 1326764
-
Detection of two loci involved in (1-->3)-beta-glucan (curdlan) biosynthesis by Agrobacterium sp. ATCC31749, and comparative sequence analysis of the putative curdlan synthase gene.
Glycobiology. 1999 Jan;9(1):31-41
PMID: 9884404
-
A 1,4-β-D-glucan-synthase system from Dictyostelium discoideum.
Planta. 1990 Feb;180(3):324-32
PMID: 24202010
-
Genetic organization of the cellulose synthase operon in Acetobacter xylinum.
Proc Natl Acad Sci U S A. 1990 Oct;87(20):8130-4
PMID: 2146681
-
Ordered yeast artificial chromosome clones representing the Dictyostelium discoideum genome.
Proc Natl Acad Sci U S A. 1996 May 28;93(11):5562-6
PMID: 8643615
-
Semimicro determination of cellulose in biological materials.
Anal Biochem. 1969 Dec;32(3):420-4
PMID: 5361396
-
The irregular xylem3 locus of Arabidopsis encodes a cellulose synthase required for secondary cell wall synthesis.
Plant Cell. 1999 May;11(5):769-80
PMID: 10330464
-
CELLULOSE BIOSYNTHESIS: Exciting Times for A Difficult Field of Study.
Annu Rev Plant Physiol Plant Mol Biol. 1999 Jun;50:245-276
PMID: 15012210
-
Cell type regulation in response to expression of ricin A in Dictyostelium.
Dev Biol. 1993 Nov;160(1):85-98
PMID: 8224551
-
Developmental signal transduction pathways uncovered by genetic suppressors.
Proc Natl Acad Sci U S A. 1996 Dec 24;93(26):15260-5
PMID: 8986798