TY - JOUR
T1 - RNA splicing and in vivo expression of the intron-containing td gene of bacteriophage T4
AU - Belfort, Marlene
AU - Pedersen-Lane, Joan
AU - Ehrenman, Karen
AU - Chu, Frederick K.
AU - Maley, Gladys F.
AU - Maley, Frank
AU - McPheeters, David S.
AU - Gold, Larry
PY - 1986
Y1 - 1986
N2 - The splice junction sequence of td mRNA from T4-infected cells has been determined (5 '4. GGU-CUA4. 3') and shown to be identical to that of the RNA ligation product encoded by the cloned gene [Belfort et al. Cell 41 (1985) 375-382]. The RNA processing functions, T4 RNA ligase, T4 polynucleotide kinase, and the host prr gene product appear not to be essential for exon ligation; neither are the host endoribonucleases RNase III, RNase P and RNase E required for intron excision. While these results are consistent with the autocatalytic splicing mechanism demonstrated in vitro [Chu et al. J. Biol. Chem. 260 (1985) 10680-10688], they leave unanswered the question of which protein(s), if any, might stimulate the in vivo reaction. Analysis of the products of the cloned td gene has led to identification of two td-encoded polypeptides, namely a polypeptide corresponding to the exon-I-coding sequence (NH2-TS), and the catalytically active thymidylate synthase (TS). Kinetic and nucleotide sequence data provide evidence that NH2-TS is the product of the primary transcript and that TS is encoded by spliced mRNA. These results suggest that splicing may provide a switch controlling the relative expression of NH2-TS and TS, two proteins with markedly different temporal appearances despite their identical transcriptional and translational start sites.
AB - The splice junction sequence of td mRNA from T4-infected cells has been determined (5 '4. GGU-CUA4. 3') and shown to be identical to that of the RNA ligation product encoded by the cloned gene [Belfort et al. Cell 41 (1985) 375-382]. The RNA processing functions, T4 RNA ligase, T4 polynucleotide kinase, and the host prr gene product appear not to be essential for exon ligation; neither are the host endoribonucleases RNase III, RNase P and RNase E required for intron excision. While these results are consistent with the autocatalytic splicing mechanism demonstrated in vitro [Chu et al. J. Biol. Chem. 260 (1985) 10680-10688], they leave unanswered the question of which protein(s), if any, might stimulate the in vivo reaction. Analysis of the products of the cloned td gene has led to identification of two td-encoded polypeptides, namely a polypeptide corresponding to the exon-I-coding sequence (NH2-TS), and the catalytically active thymidylate synthase (TS). Kinetic and nucleotide sequence data provide evidence that NH2-TS is the product of the primary transcript and that TS is encoded by spliced mRNA. These results suggest that splicing may provide a switch controlling the relative expression of NH2-TS and TS, two proteins with markedly different temporal appearances despite their identical transcriptional and translational start sites.
KW - RNA processing
KW - Splice junction sequence
KW - regulatory role for splicing
KW - thymidylate synthase
KW - two td-encoded proteins
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U2 - 10.1016/0378-1119(86)90271-4
DO - 10.1016/0378-1119(86)90271-4
M3 - Article
C2 - 2422090
AN - SCOPUS:0022449742
SN - 0378-1119
VL - 41
SP - 93
EP - 102
JO - Gene
JF - Gene
IS - 1
ER -