A putative plant homolog of the yeast beta-1,3-glucan synthase subunit FKS1 from cotton (Gossypium hirsutum L.) fibers

A novel plant gene CFL1 was cloned from cotton (Gossypium hirsutum L.) fibe rs by expressed sequence tag (EST) database searching and 5'-RACE (rapid am plification of cDNA ends). This gene shows sequence homology with FKS1 whic h has been identified as the putative catalytic subunit of the yeast beta - 1,3-glucan synthase. It encodes a protein (CFL1p) of 219 kDa with 13 deduce d transmembrane helices and 2 large hydrophilic domains, one of which is at the N-terminus and the other in the internal region of the polypeptide. CF L1 displays 21% identity and 41% similarity to FKS1 at the amino acid level over its entire length, with 31% identity and 52% similarity for the hydro philic central domain. Using RNA and protein blot analysis, CFL1 was found to be expressed at higher levels in cotton fibers during primary wall devel opment. CFL1 also had a strong expression in young roots. Using a calmoduli n (CaM)-gel overlay assay, the hydrophilic N-terminal domain of CFL1p was s hown to bind to CaM, while the hydrophilic central domain did not. A putati ve CaM-binding domain, 16 amino acids long, was predicted in the hydrophili c N-terminal domain. Moreover, a product-entrapment assay demonstrated that a protein associated with an in vitro-synthesized callose pellet could be labeled by anti-CFL1 antibodies. Our finding suggests that CFL1 is a putati ve plant homolog of the yeast beta -1,3-glucan synthase subunit FKS1 and co uld be involved in callose synthesis.