• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of molcellbPermissionsJournals.ASM.orgJournalMCB ArticleJournal InfoAuthorsReviewers
Mol Cell Biol. Apr 1991; 11(4): 1921–1926.
PMCID: PMC359875

Insertion of part of an intron into the 5' untranslated region of a Caenorhabditis elegans gene converts it into a trans-spliced gene.

Abstract

In nematodes, the RNA products of some genes are trans-spliced to a 22-nucleotide spliced leader (SL), while the RNA products of other genes are not. In Caenorhabditis elegans, there are two SLs, SL1 and SL2, donated by two distinct small nuclear ribonucleoprotein particles in a process functionally quite similar to nuclear intron removal. We demonstrate here that it is possible to convert a non-trans-spliced gene into a trans-spliced gene by placement of an intron missing only the 5' splice site into the 5' untranslated region. Stable transgenic strains were isolated expressing a gene in which 69 nucleotides of a vit-5 intron, including the 3' splice site, were inserted into the 5' untranslated region of a vit-2/vit-6 fusion gene. The RNA product of this gene was examined by primer extension and PCR amplification. Although the vit-2/vit-6 transgene product is not normally trans-spliced, the majority of transcripts from this altered gene were trans-spliced to SL1. We termed the region of a trans-spliced mRNA precursor between the 5' end and the first 3' splice site an "outron." Our results suggest that if a transcript begins with intronlike sequence followed by a 3' splice site, this alone may constitute an outron and be sufficient to demarcate a transcript as a trans-splice acceptor. These findings leave open the possibility that specific sequences are required to increase the efficiency of trans-splicing.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.5M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bektesh SL, Hirsh DI. C. elegans mRNAs acquire a spliced leader through a trans-splicing mechanism. Nucleic Acids Res. 1988 Jun 24;16(12):5692–5692. [PMC free article] [PubMed]
  • Blumenthal T, Squire M, Kirtland S, Cane J, Donegan M, Spieth J, Sharrock W. Cloning of a yolk protein gene family from Caenorhabditis elegans. J Mol Biol. 1984 Mar 25;174(1):1–18. [PubMed]
  • Blumenthal T, Thomas J. Cis and trans mRNA splicing in C. elegans. Trends Genet. 1988 Nov;4(11):305–308. [PubMed]
  • Brenner S. The genetics of Caenorhabditis elegans. Genetics. 1974 May;77(1):71–94. [PMC free article] [PubMed]
  • Bruzik JP, Steitz JA. Spliced leader RNA sequences can substitute for the essential 5' end of U1 RNA during splicing in a mammalian in vitro system. Cell. 1990 Sep 7;62(5):889–899. [PubMed]
  • Bruzik JP, Van Doren K, Hirsh D, Steitz JA. Trans splicing involves a novel form of small nuclear ribonucleoprotein particles. Nature. 1988 Oct 6;335(6190):559–562. [PubMed]
  • Chirgwin JM, Przybyla AE, MacDonald RJ, Rutter WJ. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. [PubMed]
  • Cummins C, Anderson P. Regulatory myosin light-chain genes of Caenorhabditis elegans. Mol Cell Biol. 1988 Dec;8(12):5339–5349. [PMC free article] [PubMed]
  • Fire A. Integrative transformation of Caenorhabditis elegans. EMBO J. 1986 Oct;5(10):2673–2680. [PMC free article] [PubMed]
  • Fire A, Kondo K, Waterston R. Vectors for low copy transformation of C. elegans. Nucleic Acids Res. 1990 Jul 25;18(14):4269–4270. [PMC free article] [PubMed]
  • Georgi LL, Albert PS, Riddle DL. daf-1, a C. elegans gene controlling dauer larva development, encodes a novel receptor protein kinase. Cell. 1990 May 18;61(4):635–645. [PubMed]
  • Graham RW, Jones D, Candido EP. UbiA, the major polyubiquitin locus in Caenorhabditis elegans, has unusual structural features and is constitutively expressed. Mol Cell Biol. 1989 Jan;9(1):268–277. [PMC free article] [PubMed]
  • Hamlyn PH, Browniee GG, Cheng CC, Gait MJ, Milstein C. Complete sequence of constant and 3' noncoding regions of an immunoglobulin mRNA using the dideoxynucleotide method of RNA sequencing. Cell. 1978 Nov;15(3):1067–1075. [PubMed]
  • Huang XY, Barrios LA, Vonkhorporn P, Honda S, Albertson DG, Hecht RM. Genomic organization of the glyceraldehyde-3-phosphate dehydrogenase gene family of Caenorhabditis elegans. J Mol Biol. 1989 Apr 5;206(3):411–424. [PubMed]
  • Huang XY, Hirsh D. A second trans-spliced RNA leader sequence in the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8640–8644. [PMC free article] [PubMed]
  • Karn J, Brenner S, Barnett L. Protein structural domains in the Caenorhabditis elegans unc-54 myosin heavy chain gene are not separated by introns. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4253–4257. [PMC free article] [PubMed]
  • Kramer JM, French RP, Park EC, Johnson JJ. The Caenorhabditis elegans rol-6 gene, which interacts with the sqt-1 collagen gene to determine organismal morphology, encodes a collagen. Mol Cell Biol. 1990 May;10(5):2081–2089. [PMC free article] [PubMed]
  • Krause M, Hirsh D. A trans-spliced leader sequence on actin mRNA in C. elegans. Cell. 1987 Jun 19;49(6):753–761. [PubMed]
  • Laird PW. Trans splicing in trypanosomes--archaism or adaptation? Trends Genet. 1989 Jul;5(7):204–208. [PubMed]
  • MacLeod AR, Karn J, Brenner S. Molecular analysis of the unc-54 myosin heavy-chain gene of Caenorhabditis elegans. Nature. 1981 Jun 4;291(5814):386–390. [PubMed]
  • Murphy WJ, Watkins KP, Agabian N. Identification of a novel Y branch structure as an intermediate in trypanosome mRNA processing: evidence for trans splicing. Cell. 1986 Nov 21;47(4):517–525. [PubMed]
  • Nilsen TW, Shambaugh J, Denker J, Chubb G, Faser C, Putnam L, Bennett K. Characterization and expression of a spliced leader RNA in the parasitic nematode Ascaris lumbricoides var. suum. Mol Cell Biol. 1989 Aug;9(8):3543–3547. [PMC free article] [PubMed]
  • Park YS, Kramer JM. Tandemly duplicated Caenorhabditis elegans collagen genes differ in their modes of splicing. J Mol Biol. 1990 Jan 20;211(2):395–406. [PubMed]
  • Saiki RK, Gelfand DH, Stoffel S, Scharf SJ, Higuchi R, Horn GT, Mullis KB, Erlich HA. Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science. 1988 Jan 29;239(4839):487–491. [PubMed]
  • Spieth J, Blumenthal T. The Caenorhabditis elegans vitellogenin gene family includes a gene encoding a distantly related protein. Mol Cell Biol. 1985 Oct;5(10):2495–2501. [PMC free article] [PubMed]
  • Spieth J, Denison K, Kirtland S, Cane J, Blumenthal T. The C. elegans vitellogenin genes: short sequence repeats in the promoter regions and homology to the vertebrate genes. Nucleic Acids Res. 1985 Jul 25;13(14):5283–5295. [PMC free article] [PubMed]
  • Spieth J, Denison K, Zucker E, Blumenthal T. The nucleotide sequence of a nematode vitellogenin gene. Nucleic Acids Res. 1985 Oct 11;13(19):7129–7138. [PMC free article] [PubMed]
  • Spieth J, MacMorris M, Broverman S, Greenspoon S, Blumenthal T. Regulated expression of a vitellogenin fusion gene in transgenic nematodes. Dev Biol. 1988 Nov;130(1):285–293. [PubMed]
  • Sutton RE, Boothroyd JC. Evidence for trans splicing in trypanosomes. Cell. 1986 Nov 21;47(4):527–535. [PubMed]
  • Takacs AM, Denker JA, Perrine KG, Maroney PA, Nilsen TW. A 22-nucleotide spliced leader sequence in the human parasitic nematode Brugia malayi is identical to the trans-spliced leader exon in Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1988 Nov;85(21):7932–7936. [PMC free article] [PubMed]
  • Thomas J, Lea K, Zucker-Aprison E, Blumenthal T. The spliceosomal snRNAs of Caenorhabditis elegans. Nucleic Acids Res. 1990 May 11;18(9):2633–2642. [PMC free article] [PubMed]
  • Thomas JD, Conrad RC, Blumenthal T. The C. elegans trans-spliced leader RNA is bound to Sm and has a trimethylguanosine cap. Cell. 1988 Aug 12;54(4):533–539. [PubMed]
  • Van Doren K, Hirsh D. Trans-spliced leader RNA exists as small nuclear ribonucleoprotein particles in Caenorhabditis elegans. Nature. 1988 Oct 6;335(6190):556–559. [PubMed]
  • Zeng WL, Alarcon CM, Donelson JE. Many transcribed regions of the Onchocerca volvulus genome contain the spliced leader sequence of Caenorhabditis elegans. Mol Cell Biol. 1990 Jun;10(6):2765–2773. [PMC free article] [PubMed]

Articles from Molecular and Cellular Biology are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...