Processing of the intron-containing thymidylate synthase (td) gene of phage T4 is at the RNA level

Marlene Belfort; Joan Pedersen-Lane; Deborah West; Karen Ehrenman; Gladys Maley; Frederick Chu; Frank Maley

doi:10.1016/S0092-8674(85)80010-6

Processing of the intron-containing thymidylate synthase (td) gene of phage T4 is at the RNA level

Marlene Belfort, Joan Pedersen-Lane, Deborah West, Karen Ehrenman, Gladys Maley, Frederick Chu, Frank Maley

Research output: Contribution to journal › Article › peer-review

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Abstract

The interrupted T4 phage td gene, which encodes thymidylate synthase, is the first known example of an intron-containing prokaryotic structural gene. Analysis of td-encoded transcripts provides evidence in favor of maturation at the RNA level. Northern blotting with T4 RNA and with region-specific probes revealed three classes of RNA: diffuse premessages (ca. 2.5 kb), a low-abundance mature mRNA (ca. 1.3 kb), and an abundant free intron RNA (ca. 1.0 kb). The existence of covalently joined mature mRNA was suggested by hybridization and S1 protection experiments and was confirmed by primer extension analysis of the splice junction. In analogy to expression of interrupted eukaryotic genes, these results are consistent with an RNA processing model that would account for the direct gene transcript serving as precursor for both free intron RNA and a spliced mRNA that is colinear with the thymidylate synthase product.

Original language	English (US)
Pages (from-to)	375-382
Number of pages	8
Journal	Cell
Volume	41
Issue number	2
DOIs	https://doi.org/10.1016/S0092-8674(85)80010-6
State	Published - Jun 1985
Externally published	Yes

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology

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10.1016/S0092-8674(85)80010-6

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title = "Processing of the intron-containing thymidylate synthase (td) gene of phage T4 is at the RNA level",

abstract = "The interrupted T4 phage td gene, which encodes thymidylate synthase, is the first known example of an intron-containing prokaryotic structural gene. Analysis of td-encoded transcripts provides evidence in favor of maturation at the RNA level. Northern blotting with T4 RNA and with region-specific probes revealed three classes of RNA: diffuse premessages (ca. 2.5 kb), a low-abundance mature mRNA (ca. 1.3 kb), and an abundant free intron RNA (ca. 1.0 kb). The existence of covalently joined mature mRNA was suggested by hybridization and S1 protection experiments and was confirmed by primer extension analysis of the splice junction. In analogy to expression of interrupted eukaryotic genes, these results are consistent with an RNA processing model that would account for the direct gene transcript serving as precursor for both free intron RNA and a spliced mRNA that is colinear with the thymidylate synthase product.",

author = "Marlene Belfort and Joan Pedersen-Lane and Deborah West and Karen Ehrenman and Gladys Maley and Frederick Chu and Frank Maley",

note = "Funding Information: We thank John Abelsen and Nina Piccini for their incisive suggestions in regard to the nuclease protection experiments, and David McPhee-teres and Peter Gegenheimer for helpful advice on primer extension analysis. We gratefully acknowledge Larry Gold and David McPhee-teres for communicating their splice junction sequence data to us. The support of Tom Cech is much appreciated. K. E., a graduate student in the Department of Microbiology and Immunology at Albany Medical College, is supported in part by a tuition waiver from the cotlege. This work was supported in part by grants from the National Institutes of Health (GM33314 to M.B. and GM26387 to EM.) and the National Science Foundation (PCM8118368 to M.B. and PCM8402005 to G.RM.).",

year = "1985",

month = jun,

doi = "10.1016/S0092-8674(85)80010-6",

language = "English (US)",

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T1 - Processing of the intron-containing thymidylate synthase (td) gene of phage T4 is at the RNA level

AU - Belfort, Marlene

AU - Pedersen-Lane, Joan

AU - West, Deborah

AU - Ehrenman, Karen

AU - Maley, Gladys

AU - Chu, Frederick

AU - Maley, Frank

N1 - Funding Information: We thank John Abelsen and Nina Piccini for their incisive suggestions in regard to the nuclease protection experiments, and David McPhee-teres and Peter Gegenheimer for helpful advice on primer extension analysis. We gratefully acknowledge Larry Gold and David McPhee-teres for communicating their splice junction sequence data to us. The support of Tom Cech is much appreciated. K. E., a graduate student in the Department of Microbiology and Immunology at Albany Medical College, is supported in part by a tuition waiver from the cotlege. This work was supported in part by grants from the National Institutes of Health (GM33314 to M.B. and GM26387 to EM.) and the National Science Foundation (PCM8118368 to M.B. and PCM8402005 to G.RM.).

PY - 1985/6

Y1 - 1985/6

N2 - The interrupted T4 phage td gene, which encodes thymidylate synthase, is the first known example of an intron-containing prokaryotic structural gene. Analysis of td-encoded transcripts provides evidence in favor of maturation at the RNA level. Northern blotting with T4 RNA and with region-specific probes revealed three classes of RNA: diffuse premessages (ca. 2.5 kb), a low-abundance mature mRNA (ca. 1.3 kb), and an abundant free intron RNA (ca. 1.0 kb). The existence of covalently joined mature mRNA was suggested by hybridization and S1 protection experiments and was confirmed by primer extension analysis of the splice junction. In analogy to expression of interrupted eukaryotic genes, these results are consistent with an RNA processing model that would account for the direct gene transcript serving as precursor for both free intron RNA and a spliced mRNA that is colinear with the thymidylate synthase product.

AB - The interrupted T4 phage td gene, which encodes thymidylate synthase, is the first known example of an intron-containing prokaryotic structural gene. Analysis of td-encoded transcripts provides evidence in favor of maturation at the RNA level. Northern blotting with T4 RNA and with region-specific probes revealed three classes of RNA: diffuse premessages (ca. 2.5 kb), a low-abundance mature mRNA (ca. 1.3 kb), and an abundant free intron RNA (ca. 1.0 kb). The existence of covalently joined mature mRNA was suggested by hybridization and S1 protection experiments and was confirmed by primer extension analysis of the splice junction. In analogy to expression of interrupted eukaryotic genes, these results are consistent with an RNA processing model that would account for the direct gene transcript serving as precursor for both free intron RNA and a spliced mRNA that is colinear with the thymidylate synthase product.

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Processing of the intron-containing thymidylate synthase (td) gene of phage T4 is at the RNA level

Abstract

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