A localisation signal in the 3' untranslated region of c-myc mRNA targets c-myc mRNA and beta-globin reporter sequences to the perinuclear cytoplasm and cytoskeletal-bound polysomes.
There is increasing evidence that in mammalian cells some mRNAs are localised to specific parts of the cytoplasm and a proportion of mRNAs and polyribosomes are associated with the cytoskeleton. It has been shown previously that c-myc mRNA is present in the perinuclear cytoplasm and associated with the cytoskeleton, and that this localisation is dependent upon the 3' untranslated region of the mRNA. The present studies show that in transfected fibroblasts the c-myc 3' untranslated region is able to localise beta-globin reporter sequences to the perinuclear cytoplasm. Studies with constructs containing deletions within the 3' untranslated region identify the region between bases 194 and 280 as critical for localisation. Transfection of cells with constructs in which this region is linked to beta-globin sequences showed that it was sufficient to localise the chimaeric transcripts to the perinuclear cytoplasm and to cytoskeletal-bound polyribosomes. Transfection with constructs containing a mutated AUUUA sequence within the 194-280 base region showed that this conserved AUUUA is required for targeting of both c-myc mRNA and a chimaeric transcript of beta-globin transcripts linked to the c-myc 3' untranslated region. The region between bases 194 and 280 did not induce instability of beta-globin transcripts and the AUUUA mutation had little effect upon mRNA stability. We propose that this 86 nt region of the 3' untranslated region contains a localisation signal to target c-myc mRNA so that it is retained on cytoskeletal-bound polysomes in the perinuclear cytoplasm; a conserved AUUUA sequence appears to be a critical part of this signal.
Veyrune
J L
JL
Institut de Genetique Moleculaire de Montpellier, UMR 5535, CNRS, Montpellier, France.
Campbell
G P
GP
Wiseman
J
J
Blanchard
J M
JM
Hesketh
J E
JE
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D017930:Genes, Reporter; D016259:Genes, myc; D005914:Globins / Q000235:genetics; D017403:In Situ Hybridization; D009154:Mutation; D011132:Polyribosomes / Q000378:metabolism; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism*; D051381:Rats; D017384:Sequence Deletion; D015398:Signal Transduction; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection
D012333:RNA, Messenger; D005914:Globins