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Localization of sequences required for size-specific splicing of a small Drosophila intron in vitro.
J Mol Biol. 1995 Oct 27; 253(3):426-37.JM

Abstract

Many introns in Drosophila and other invertebrates are less than 80 nucleotides in length, too small to be recognized by the vertebrate splicing machinery. Comparison of nuclear splicing extracts from human HeLa and Drosophila Kc cells has revealed species-specificity, consistent with the observed size differences. Here we present additional results with the 68 nucleotide fifth intron of the Drosophila myosin heavy chain gene. As observed with the 74 nucleotide second intron of the Drosophila white gene, the wild-type myosin intron is accurately spliced in a homologous extract, and increasing the size by 16 nucleotides both eliminates splicing in the Drosophila extract and allows accurate splicing in the human extract. In contrast to previous results, however, an upstream cryptic 5' splice site is activated when the wild-type myosin intron is tested in a human HeLa cell nuclear extract, resulting in the removal of a 98 nucleotide intron. The size dependence of splicing in Drosophila extracts is also intron-specific; we noted that a naturally larger (150 nucleotide) intron from the ftz gene is efficiently spliced in Kc cell extracts that do not splice enlarged introns (of 84, 90, 150 or 350 nucleotides) derived from the 74 nucleotide white intron. Here, we have exploited that observation, using a series of hybrid introns to show that a region of 46 nucleotides at the 3' end of the white intron is sufficient to confer the species-specific size effect. At least two sequence elements within this region, yet distinct from previously described branchpoint and pyrimidine tract signals, are required for efficient splicing of small hybrid introns in vitro.

Authors+Show Affiliations

Department of Biological Sciences, Columbia University, New York, NY 10027, USA.No affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

7473725

Citation

Guo, M, and S M. Mount. "Localization of Sequences Required for Size-specific Splicing of a Small Drosophila Intron in Vitro." Journal of Molecular Biology, vol. 253, no. 3, 1995, pp. 426-37.
Guo M, Mount SM. Localization of sequences required for size-specific splicing of a small Drosophila intron in vitro. J Mol Biol. 1995;253(3):426-37.
Guo, M., & Mount, S. M. (1995). Localization of sequences required for size-specific splicing of a small Drosophila intron in vitro. Journal of Molecular Biology, 253(3), 426-37.
Guo M, Mount SM. Localization of Sequences Required for Size-specific Splicing of a Small Drosophila Intron in Vitro. J Mol Biol. 1995 Oct 27;253(3):426-37. PubMed PMID: 7473725.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Localization of sequences required for size-specific splicing of a small Drosophila intron in vitro. AU - Guo,M, AU - Mount,S M, PY - 1995/10/27/pubmed PY - 1995/10/27/medline PY - 1995/10/27/entrez SP - 426 EP - 37 JF - Journal of molecular biology JO - J Mol Biol VL - 253 IS - 3 N2 - Many introns in Drosophila and other invertebrates are less than 80 nucleotides in length, too small to be recognized by the vertebrate splicing machinery. Comparison of nuclear splicing extracts from human HeLa and Drosophila Kc cells has revealed species-specificity, consistent with the observed size differences. Here we present additional results with the 68 nucleotide fifth intron of the Drosophila myosin heavy chain gene. As observed with the 74 nucleotide second intron of the Drosophila white gene, the wild-type myosin intron is accurately spliced in a homologous extract, and increasing the size by 16 nucleotides both eliminates splicing in the Drosophila extract and allows accurate splicing in the human extract. In contrast to previous results, however, an upstream cryptic 5' splice site is activated when the wild-type myosin intron is tested in a human HeLa cell nuclear extract, resulting in the removal of a 98 nucleotide intron. The size dependence of splicing in Drosophila extracts is also intron-specific; we noted that a naturally larger (150 nucleotide) intron from the ftz gene is efficiently spliced in Kc cell extracts that do not splice enlarged introns (of 84, 90, 150 or 350 nucleotides) derived from the 74 nucleotide white intron. Here, we have exploited that observation, using a series of hybrid introns to show that a region of 46 nucleotides at the 3' end of the white intron is sufficient to confer the species-specific size effect. At least two sequence elements within this region, yet distinct from previously described branchpoint and pyrimidine tract signals, are required for efficient splicing of small hybrid introns in vitro. SN - 0022-2836 UR - https://www.unboundmedicine.com/medline/citation/7473725/Localization_of_sequences_required_for_size_specific_splicing_of_a_small_Drosophila_intron_in_vitro_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0022-2836(85)70564-5 DB - PRIME DP - Unbound Medicine ER -