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Nucleic Acids Research, 1984, Vol. 12, No. 22 8363-8380
© 1984

MOLECULAR BIOLOGY

Subcellular localization of RNAs in transfected cells: role of sequences at the 5' terminus

Aixa Alfonso-Pizarro, David P. Carlson and Jeffrey Ross

McArdle Laboratory for Cancer Research 450 North Randall Avenue, Madison, WI 53706, USA

Received August 10, 1984. Revised October 22, 1984. Accepted October 22, 1984.

We have investigated the intracellular location of RNAs transcribed from transfected DNA. COS cells transfected with a clone containing the human adult ß globin gene contain three classes of globin RNAs. Their 3' termini and splice sites are indistinguishable from those of mature reticulocyte ß globin mRNA, and they are polyadenylated. However, as determined by S1 mapping, their 5' sequences are different. The 5' terminus of one is the same as that of mature ß globin mRNA (+1, cap site). The presumed 5' terminus of the second is located 30 nucleotides downstream from the cap site (+30). The third class contains additional nucleotides transcribed from sequences located 5' to the cap site (5' upstream RNA). The 5' upstream RNA molecules are restricted to the nucleus and are more stable than heterogeneous nuclear RNA. The +30 and +1 RNAs are located primarily in the cytoplasm. The data support the notion that nucleotide sequences and/or secondary modifications in the 5' region determine if an RNA is to be transported.


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