A novel fatty Acyl-CoA Synthetase is required for pollen development and sporopollenin biosynthesis in Arabidopsis.
Acyl-CoA Synthetase (ACOS) genes are related to 4-coumarate:CoA ligase (4CL) but have distinct functions. The Arabidopsis thaliana ACOS5 protein is in clade A of Arabidopsis ACOS proteins, the clade most closely related to 4CL proteins. This clade contains putative nonperoxisomal ACOS enzymes conserved in several angiosperm lineages and in the moss Physcomitrella patens. Although its function is unknown, ACOS5 is preferentially expressed in the flowers of all angiosperms examined. Here, we show that an acos5 mutant produced no pollen in mature anthers and no seeds by self-fertilization and was severely compromised in pollen wall formation apparently lacking sporopollenin or exine. The phenotype was first evident at stage 8 of anther development and correlated with maximum ACOS5 mRNA accumulation in tapetal cells at stages 7 to 8. Green fluorescent protein-ACOS5 fusions showed that ACOS5 is located in the cytoplasm. Recombinant ACOS5 enzyme was active against oleic acid, allowing kinetic constants for ACOS5 substrates to be established. Substrate competition assays indicated broad in vitro preference of the enzyme for medium-chain fatty acids. We propose that ACOS5 encodes an enzyme that participates in a conserved and ancient biochemical pathway required for sporopollenin monomer biosynthesis that may also include the Arabidopsis CYP703A2 and MS2 enzymes.
de Azevedo Souza
Clarice
C
Department of Botany, University of British Columbia, Vancouver, BC V6T 1Z4, Canada.
Kim
Sung Soo
SS
Koch
Stefanie
S
Kienow
Lucie
L
Schneider
Katja
K
McKim
Sarah M
SM
Haughn
George W
GW
Kombrink
Erich
E
Douglas
Carl J
CJ
D000483:Alleles; D017360:Arabidopsis / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D001667:Binding, Competitive; D001704:Biopolymers / Q000096:biosynthesis* / Q000737:chemistry; D002338:Carotenoids / Q000096:biosynthesis* / Q000737:chemistry; D003066:Coenzyme A Ligases / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D003593:Cytoplasm / Q000378:metabolism; D035264:Flowers / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000032:analysis; D007700:Kinetics; D009154:Mutation; D010802:Phylogeny; D011058:Pollen / Q000254:growth & development* / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000032:analysis; D013379:Substrate Specificity
D029681:Arabidopsis Proteins; D001704:Biopolymers; D011993:Recombinant Fusion Proteins; C009800:sporopollenin; D049452:Green Fluorescent Proteins; D002338:Carotenoids; C542001:ACOS5 protein, Arabidopsis; D003066:Coenzyme A Ligases