PRH75, a new nucleus-localized member of the DEAD-box protein family from higher plants.
The putative RNA helicases of the DEAD-box protein family are involved in pre-mRNA splicing, rRNA maturation, ribosome assembly, and translation. Members of this protein family have been identified in organisms from Escherichia coli to humans, but except for the translation initiation factor 4A, there have been no reports on the characterization of other DEAD-box proteins from plants. Here we report on a novel member of the DEAD-box protein family, the plant RNA helicase 75 (PRH75). PRH75 is localized in the nucleus and contains two domains for RNA binding. One is located at the C terminus and is similar to RGG RNA-binding domains of nucleus-localized RNA-binding proteins. The other one is located between amino acids 308 and 622, a region containing the conserved motif VI characteristic of DEAD-box proteins and known as the RNA-binding site of eIF-4A. The N-terminal 81 amino acids are sufficient for nuclear targeting of the protein. Northern and Western blot analyses show that PRH75 is mainly expressed in young and rapidly developing tissues. The purified recombinant PRH75 has a weak ATPase activity which is barely stimulated by RNA ligands. The fractionation of spinach whole-cell extracts by glycerol gradient centrifugation and gel filtration on a Superdex 200 column shows that the protein exists in a complex of about 500 kDa. Possible biological functions of PRH75 as well as structure-function relationships in the context of its modular primary structure are discussed.
Lorković
Z J
ZJ
Botanisches Institut der Ludwig-Maximilians-Universität, Munich, Germany. oel@botanik.biologie.uni-muenchen.de
Herrmann
R G
RG
Oelmüller
R
R
D000595:Amino Acid Sequence; D029681:Arabidopsis Proteins; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D053487:DEAD-box RNA Helicases; D018076:DNA, Complementary / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D017343:Genes, Plant; D006801:Humans; D008969:Molecular Sequence Data; D015394:Molecular Structure; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020365:RNA Helicases; D012316:RNA Nucleotidyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D018749:RNA, Plant / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D018724:Spinacia oleracea / Q000235:genetics / Q000254:growth & development / Q000378:metabolism
D029681:Arabidopsis Proteins; D018076:DNA, Complementary; D018744:DNA, Plant; D009687:Nuclear Proteins; D010940:Plant Proteins; D018749:RNA, Plant; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; C105180:PRH75 protein, Arabidopsis; D012316:RNA Nucleotidyltransferases; D053487:DEAD-box RNA Helicases; D020365:RNA Helicases