A nodule-specific gene encoding a subtilisin-like protease is expressed in early stages of actinorhizal nodule development.
To identify genes specifically expressed during early stages of actinorhizal nodule development, a cDNA library made from poly(A) RNA from root nodules of Alnus glutinosa was screened differentially with nodule and root cDNA, respectively. Seven nodule-enhanced and four nodule-specific cDNA clones were isolated. By using in situ hybridization, two of the nodule-specific cDNAs were shown to be expressed at the highest levels in infected cells before the onset of nitrogen fixation; one of them, ag12 (A. glutinosa), was examined in detail. Sequencing showed that ag12 codes for a serine protease of the subtilisin (EC 3.4.21.14) family. Subtilisins previously appeared to be limited to microorganisms. However, subtilisin-like serine proteases have recently been found in archaebacteria, fungi, and yeasts as well as in mammals; a plant subtilisin has also been sequenced. In yeast and mammals, subtilases are responsible for processing peptide hormones. A homolog of ag12, ara12, was identified in Arabidopsis; it was expressed in all organs, and its expression levels were highest during silique development. Hence, our study shows that subtilases are also involved in both symbiotic and nonsymbiotic processes in plant development.
Ribeiro
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A
Department of Molecular Biology, Agricultural University, Wageningen, The Netherlands.
Akkermans
A D
AD
van Kammen
A
A
Bisseling
T
T
Pawlowski
K
K
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics; D029681:Arabidopsis Proteins; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D003904:Mitosporic Fungi; D008969:Molecular Sequence Data; D009586:Nitrogen Fixation; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D018517:Plant Roots / Q000201:enzymology* / Q000254:growth & development / Q000382:microbiology; D017386:Sequence Homology, Amino Acid; D013381:Subtilisins / Q000235:genetics* / Q000378:metabolism; D013559:Symbiosis; D014197:Trees
D029681:Arabidopsis Proteins; D010940:Plant Proteins; D013381:Subtilisins; C095312:ag12 protein, Alnus glutinosa; C095313:ara12 protein, Arabidopsis