Molecular cloning and characterisation of a maize cDNA for a homologue of the large subunit of the eukaryotic initiation factor 3 (eIF3).
In order to identify genes that are specifically expressed in distinct cell populations of the maize root apex, we have constructed PCR-directed cDNA libraries from microdissected populations of cells, and screened them by differential hybridisation. A meristem-specific cDNA was isolated and characterised. This cDNA, termed ZmeIF3A, encodes a protein homologous to the large subunit of the eukaryotic translation Initiation Factor 3 (eIF3), which is an essential multi-protein complex for the initiation of protein synthesis. The ZmeIF3A protein is most similar to the yeast homologue RPG1, lacking the repeated C-terminal domain characteristic of its mammalian counterparts. However, despite this similarity, it fails to replace the RPG1 protein in complementation experiments on yeast mutants. Analysis of gene expression in situ showed that the ZmeIF3A transcript is expressed in the region of the root meristem surrounding the central stele. ZmeIF3A mRNA is also expressed in the young root, the male inflorescence, and the developing cob and seed. In maize, ZmeIF3A is encoded by one or two genomic sequences. This is the first report on the isolation and characterisation of a cDNA from higher plants that encodes a product homologous to a component of the eIF3 complex.
Institute of Molecular and Cell Biology, National University of Singapore, Republic of Singapore. paolosabelli@iol.it
P A
PA
Sabelli
S R
SR
Burgess
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L
Valásek
P R
PR
Shewry
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary; D039621:Eukaryotic Initiation Factor-3; D046911:Macromolecular Substances; D008322:Mammals; D018519:Meristem; D008969:Molecular Sequence Data; D010448:Peptide Initiation Factors / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D018517:Plant Roots; D016133:Polymerase Chain Reaction; D017433:Protein Structure, Secondary; D012639:Seeds / Q000502:physiology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D003313:Zea mays / Q000235:genetics* / Q000502:physiology
D017931:DNA Primers; D018076:DNA, Complementary; D039621:Eukaryotic Initiation Factor-3; D046911:Macromolecular Substances; D010448:Peptide Initiation Factors