The rice [Oryza sativa] nucellin gene ortholog OsAsp1 encodes an active aspartic protease without a plant-specific insert and is strongly expressed in early embryo

The barley nucellin gene was reported to be nucellus specific in its expression and was hypothesized to play a role in the programmed cell death of the nucellus as an aspartic protease. Here we provide direct evidence that the rice ortholog encodes an active aspartic protease, but we prefer the name...

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Bibliographic Details
Published in:Plant and cell physiology 2005-01, Vol.46 (1), p.87-98
Main Authors: Bi, X.(International Rice Research Inst., Manila (Philippines)), Khush, G.S, Bennett, J
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Arabidopsis
Aspartic Acid Endopeptidases - genetics
Base Sequence
Chromosome Mapping
Chromosomes, Plant - genetics
Cloning, Molecular
DAF
days after flowering
DESARROLLO EMBRIONARIO
DEVELOPPEMENT EMBRYONNAIRE
DIG
digoxygenin
DNA, Plant - genetics
double haploid
EMBRYONIC DEVELOPMENT
endoplasmic reticulum
Escherichia coli
EST
EXPRESION GENICA
expressed sequence tag
EXPRESSION DES GENES
fluorescein thiocarbamoyl
FTC
GENE
GENE EXPRESSION
Gene Expression Regulation, Developmental
Gene Expression Regulation, Enzymologic
Gene Expression Regulation, Plant
GENES
Genes, Plant
Hordeum - enzymology
Hordeum - genetics
HORDEUM VULGARE
IPTG
isopropyl 1-thio-β-d-galactopyranoside
Keywords: Aspartic protease — Embryo — Gene expression — Recombinant protein — Rice (Oryza sativa L.)
MALDI-TOF
matrix-assisted laser desorption/ionization time of flight
Molecular Sequence Data
Oryza - embryology
Oryza - enzymology
Oryza - genetics
ORYZA SATIVA
PCD
Peptide Mapping
peroxisome targeting sequence
Phylogeny
Plant Proteins - genetics
plant-specific insert
programmed cell death
PROTEASAS
PROTEASE
PROTEASES
PSI
PTS
Recombinant Proteins - genetics
reverse transcriptase–polymerase chain reaction
RNA, Messenger - genetics
RNA, Messenger - metabolism
RNA, Plant - genetics
RNA, Plant - metabolism
RT–PCR
SDS-PAGE
Sequence Homology, Amino Acid
untranslated region
UTR
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Summary:The barley nucellin gene was reported to be nucellus specific in its expression and was hypothesized to play a role in the programmed cell death of the nucellus as an aspartic protease. Here we provide direct evidence that the rice ortholog encodes an active aspartic protease, but we prefer the name aspartic proteasel (OsAsp1) to nucellin after a detailed analysis of its expression pattern in rice and barley. Northern blots. RT-PCR and RNA in situ hybridization showed that OsAsp1 is expressed most abundantly in zygotic embryos 1-2 d after fertilization. It is also expressed in pollen, nucellus, ovary wall, shoot and root meristem, coleoptiles of immature seeds, and somatic embryos. A parallel study in barley showed that the barley nucellin gene was expressed not only in the nucellus but also strong]y in embryos. Recombinant protein proOsAspl expressed in the bacterium Escherichia coli refolded and autolysed at acidic pH 3.5 in vitro, and the mature peptide displayed protease activity. Nucellin has three close homologs in rice on chromosomes 11 and 12 and in Arabidopsis on chromosomes 1 and 4. They lack the plant-specific insert that distinguishes the typical plant aspartic protease from aspartic proteases of other organisms. They constitute a new class of aspartic protease that is present in both monocots and dicots but whose function remains to be explored further.
ISSN:0032-0781
1471-9053
DOI:10.1093/pcp/pci002