Recognition and long-range interactions of a minimal nanos RNA localization signal element.
Localization of nanos (nos) mRNA to the germ plasm at the posterior pole of the Drosophila embryo is essential to activate nos translation and thereby generate abdominal segments. nos RNA localization is mediated by a large cis-acting localization signal composed of multiple, partially redundant elements within the nos 3' untranslated region. We identify a protein of approximately 75 kDa (p75) that interacts specifically with the nos +2' localization signal element. We show that the function of this element can be delimited to a 41 nucleotide domain that is conserved between D. melanogaster and D. virilis, and confers near wild-type localization when present in three copies. Two small mutations within this domain eliminate both +2' element localization function and p75 binding, consistent with a role for p75 in nos RNA localization. In the intact localization signal, the +2' element collaborates with adjacent localization elements. We show that different +2' element mutations not only abolish collaboration between the +2' and adjacent +1 element but also produce long-range deleterious effects on localization signal function. Our results suggest that higher order structural interactions within the localization signal, which requires factors such as p75, are necessary for association of nos mRNA with the germ plasm.
Bergsten
S E
SE
Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA.
Huang
T
T
Chatterjee
S
S
Gavis
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ER
D020413:3' Untranslated Regions / Q000235:genetics; D000005:Abdomen / Q000196:embryology; D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D001692:Biological Transport; D019521:Body Patterning; D017124:Conserved Sequence; D003593:Cytoplasm / Q000378:metabolism; D004330:Drosophila / Q000166:cytology / Q000196:embryology* / Q000235:genetics*; D029721:Drosophila Proteins; D005260:Female; D018507:Gene Expression Regulation, Developmental; D017403:In Situ Hybridization; D007301:Insect Hormones / Q000235:genetics; D019476:Insect Proteins / Q000235:genetics*; D008970:Molecular Weight; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D014176:Protein Biosynthesis; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*
D020413:3' Untranslated Regions; D029721:Drosophila Proteins; D007301:Insect Hormones; D019476:Insect Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; C070574:nos protein, Drosophila