Identification of Cis-acting Intron and Exon Regions in Influenza Virus NS1 MRNA That Inhibit Splicing and Cause the Formation of Aberrantly Sedimenting Presplicing Complexes

Overview

Journal Mol Cell Biol

Specialty Cell Biology

Date 1992 Mar 1

PMID 1532050

Citations 26

Authors

M E Nemeroff

U Utans

A Kramer

R M KRUG

Affiliations

Soon will be listed here.

Abstract

In in vitro splicing reactions, influenza virus NS1 mRNA was not detectably spliced, but nonetheless very efficiently formed ATP-dependent 55S complexes containing the U1, U2, U4, U5, and U6 small nuclear ribonucleoproteins (snRNPs) (C. H. Agris, M. E. Nemeroff, and R. M. Krug, Mol. Cell. Biol. 9:259-267, 1989). We demonstrate that the block in splicing was caused by two regions in NS1 mRNA: (i) a large intron region (not including the branchpoint sequence) and (ii) an 85-nucleotide 3' exon region near the 3' end of the exon. After removal of both of these regions, the 5' and 3' splice sites and branchpoint of NS1 mRNA functioned efficiently in splicing, indicating that they were not defective. The two inhibitory regions shared one property: splicing inhibition was independent of the identity of the nucleotide sequence in either region. In other respects, however, the two inhibitory regions differed. The inhibitory activity of the intron region was proportional to its length, indicating that the inhibition was probably due to size only. In contrast, the 3' exon, which was of small size, was a context element; i.e., it functioned only when it was located at a specific position in the 3' exon of NS1 mRNA. To determine how these intron and exon regions inhibited splicing, we compared the types of splicing complexes formed by intact NS1 mRNA with those formed by spliceable NS1 mRNA lacking the intron and exon regions. Splicing complexes were formed by using purified splicing factors.(ABSTRACT TRUNCATED AT 250 WORDS)

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References

KRUG R . Regulation of the extent of splicing of influenza virus NS1 mRNA: role of the rates of splicing and of the nucleocytoplasmic transport of NS1 mRNA. Mol Cell Biol. 1991; 11(2):1092-8. PMC: 359785. DOI: 10.1128/mcb.11.2.1092-1098.1991. View

Lamb R, Lai C . Sequence of interrupted and uninterrupted mRNAs and cloned DNA coding for the two overlapping nonstructural proteins of influenza virus. Cell. 1980; 21(2):475-85. DOI: 10.1016/0092-8674(80)90484-5. View

Siebel C, Rio D . Regulated splicing of the Drosophila P transposable element third intron in vitro: somatic repression. Science. 1990; 248(4960):1200-8. DOI: 10.1126/science.2161558. View

Krainer A, Conway G, Kozak D . The essential pre-mRNA splicing factor SF2 influences 5' splice site selection by activating proximal sites. Cell. 1990; 62(1):35-42. DOI: 10.1016/0092-8674(90)90237-9. View

Luytjes W, Krystal M, ENAMI M, Parvin J, Palese P . Amplification, expression, and packaging of foreign gene by influenza virus. Cell. 1989; 59(6):1107-13. DOI: 10.1016/0092-8674(89)90766-6. View

Arrigo S, Beemon K . Regulation of Rous sarcoma virus RNA splicing and stability. Mol Cell Biol. 1988; 8(11):4858-67. PMC: 365579. DOI: 10.1128/mcb.8.11.4858-4867.1988. View

Bindereif A, Green M . Ribonucleoprotein complex formation during pre-mRNA splicing in vitro. Mol Cell Biol. 1986; 6(7):2582-92. PMC: 367814. DOI: 10.1128/mcb.6.7.2582-2592.1986. View

Black D, Chabot B, Steitz J . U2 as well as U1 small nuclear ribonucleoproteins are involved in premessenger RNA splicing. Cell. 1985; 42(3):737-50. DOI: 10.1016/0092-8674(85)90270-3. View

Frendewey D, Keller W . Stepwise assembly of a pre-mRNA splicing complex requires U-snRNPs and specific intron sequences. Cell. 1985; 42(1):355-67. DOI: 10.1016/s0092-8674(85)80131-8. View

10.

Plotch S, KRUG R . In vitro splicing of influenza viral NS1 mRNA and NS1-beta-globin chimeras: possible mechanisms for the control of viral mRNA splicing. Proc Natl Acad Sci U S A. 1986; 83(15):5444-8. PMC: 386303. DOI: 10.1073/pnas.83.15.5444. View

11.

Shapiro G, Gurney Jr T, KRUG R . Influenza virus gene expression: control mechanisms at early and late times of infection and nuclear-cytoplasmic transport of virus-specific RNAs. J Virol. 1987; 61(3):764-73. PMC: 254018. DOI: 10.1128/JVI.61.3.764-773.1987. View

12.

Wieringa B, Hofer E, Weissmann C . A minimal intron length but no specific internal sequence is required for splicing the large rabbit beta-globin intron. Cell. 1984; 37(3):915-25. DOI: 10.1016/0092-8674(84)90426-4. View

13.

Lamb R, Lai C . Expression of unspliced NS1 mRNA, spliced NS2 mRNA, and a spliced chimera mRNA from cloned influenza virus NS DNA in an SV40 vector. Virology. 1984; 135(1):139-47. DOI: 10.1016/0042-6822(84)90124-7. View

14.

Lamb R, Lai C, Choppin P . Sequences of mRNAs derived from genome RNA segment 7 of influenza virus: colinear and interrupted mRNAs code for overlapping proteins. Proc Natl Acad Sci U S A. 1981; 78(7):4170-4. PMC: 319750. DOI: 10.1073/pnas.78.7.4170. View

15.

Kramer A, Utans U . Three protein factors (SF1, SF3 and U2AF) function in pre-splicing complex formation in addition to snRNPs. EMBO J. 1991; 10(6):1503-9. PMC: 452814. DOI: 10.1002/j.1460-2075.1991.tb07670.x. View

16.

Fu X, KATZ R, Skalka A, Maniatis T . The role of branchpoint and 3'-exon sequences in the control of balanced splicing of avian retrovirus RNA. Genes Dev. 1991; 5(2):211-20. DOI: 10.1101/gad.5.2.211. View

17.

KATZ R, Skalka A . Control of retroviral RNA splicing through maintenance of suboptimal processing signals. Mol Cell Biol. 1990; 10(2):696-704. PMC: 360868. DOI: 10.1128/mcb.10.2.696-704.1990. View

18.

Ge H, Manley J . A protein factor, ASF, controls cell-specific alternative splicing of SV40 early pre-mRNA in vitro. Cell. 1990; 62(1):25-34. DOI: 10.1016/0092-8674(90)90236-8. View

19.

REED R, Maniatis T . A role for exon sequences and splice-site proximity in splice-site selection. Cell. 1986; 46(5):681-90. DOI: 10.1016/0092-8674(86)90343-0. View

20.

Agris C, Nemeroff M, KRUG R . A block in mammalian splicing occurring after formation of large complexes containing U1, U2, U4, U5, and U6 small nuclear ribonucleoproteins. Mol Cell Biol. 1989; 9(1):259-67. PMC: 362168. DOI: 10.1128/mcb.9.1.259-267.1989. View