Cloning and characterization of a naturally occurring antisense RNA to human thymidylate synthase mRNA.
Based upon reverse transcription and polymerase chain reaction results with human KB cell RNA, a cDNA (i.e., 3'rTS1, 1557 nt) with complementarity to thymidylate synthase mRNA was cloned and sequenced. Northern blot analysis showed that 3'rTS1 corresponded to a cytoplasmic 1.8 kb RNA found in several tumor cell lines. The remaining 5'region of this antisense RNA was cloned by a RACE (Rapid Amplification of cDNA Ends) procedure. A full length cDNA (i.e., rTS, 1811 nt) was generated by splicing 3'rTS1 with RACE-generated cDNA. rTS RNA is likely a mRNA that contains four open reading frames. Based upon sequence analysis of the RACE cDNAs and the rTS cDNA, rTS RNA is likely processed from a gene containing at least six introns. Northern blot analysis indicates rTS RNA is expressed in a variety of human tumor cell lines and an aberrant from is expressed in a methotrexate-cell line.
Department of Experimental Therapeutics, Roswell Park Cancer Institute, Buffalo, NY 14263.
B J
BJ
Dolnick
D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D004247:DNA; D006801:Humans; D006836:Hydro-Lyases; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000235:genetics; D012322:RNA Precursors / Q000378:metabolism; D016372:RNA, Antisense / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D013940:Thymidylate Synthase / Q000235:genetics* / Q000378:metabolism; D014407:Tumor Cells, Cultured
D011506:Proteins; D012322:RNA Precursors; D016372:RNA, Antisense; D012333:RNA, Messenger; D004247:DNA; D013940:Thymidylate Synthase; C108693:ENOSF1 protein, human; D006836:Hydro-Lyases