Regulation of N-myc gene expression: use of an adenovirus vector to demonstrate posttranscriptional control.
We present evidence that differences in the levels of N-myc mRNA among different cell types are the result of posttranscriptional control. First, we noted that while steady-state mouse N-myc mRNA could be detected only in fetal mouse brain, it was transcribed at an equivalent rate in adult brain, liver, spleen, and placenta and in fetal brain. Similarly, the human N-myc gene was transcribed at an equivalent rate in HeLa cells, which do not accumulate this RNA in the cytoplasm, and cell lines G401 (a Wilms tumor-derived cell line) and SKNMc (established from a primitive neuroepithelioma), which do express N-myc RNA. As expected, the N-myc promoter functioned at equivalent rates, as demonstrated by the level of a reporter gene, when introduced into these cell types by using a recombinant adenovirus vector. The suggestion that posttranscriptional mechanisms control the level of this RNA was supported by the observation that sequences in the N-myc third exon specifically decreased the level of E1A mRNA when these sequences were placed downstream of the E1A promoter in a recombinant adenovirus. Finally, we further localized these sequences to a 600-bp fragment of the third exon by introducing various subclones of this sequence downstream of the E1A promoter in both viral and plasmid vectors.
Babiss
L E
LE
Laboratory of Molecular Cell Biology, Rockefeller University, New York, New York 10021-6399.
Friedman
J M
JM
D017456:Adenovirus Early Proteins; D000818:Animals; D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics; D005091:Exons; D015972:Gene Expression Regulation, Neoplastic; D016259:Genes, myc; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D051379:Mice; D009447:Neuroblastoma; D009856:Oncogene Proteins, Viral / Q000235:genetics; D010957:Plasmids; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection
D017456:Adenovirus Early Proteins; D004279:DNA, Viral; D009856:Oncogene Proteins, Viral; D012333:RNA, Messenger