1887

Journal of General Virology header logo

Volume 75, Issue 10

Cruciform structure of a DNA motif of parvovirus minute virus of mice (prototype strain) involved in the attenuation of gene expression Free

Abstract

It has previously been reported that the region between nucleotides 259 and 383 immediately downstream from the P4 early promoter of parvovirus minute virus of mice, prototype strain (MVMp) is responsible for transcriptional attenuation. Attenuation results from the premature pausing of RNA polymerase II within this sequence (designated to as ) and seems to depend on potential RNA secondary structure. To assess the attenuation capacity of under near physiological conditions, the early transcription unit of MVMp was replaced by the chloramphenicol acetyltransferase reporter gene under control of the early P4 promoter, in the presence or absence of . The resulting recombinant vectors were encapsidated in parvovirus particles and replicated in cells after co-infection with the wild-type virus. The fragment reduced the rate of expression of the reporter gene by approximately threefold, confirming previously reported data from transfection experiments performed in the same cellular system. This attenuation factor is unexpectedly high, considering that the ‘readthrough’ fold of the nascent viral transcript is thermodynamically more stable than the ‘attenuation’ configuration. In an attempt to elucidate this point, we sought for the presence of secondary structures in the template DNA molecule. nuclease probing of viral dsDNA revealed that the fragment had a cruciform configuration with both complementary strands folding into the computer- predicted stem-loop ‘attenuation’ structure. These observations lead us to propose that the secondary structure of the DNA template may prompt the formation of the ‘attenuation’ stem-loop in nascent mRNAs by bringing corresponding self-complementary sequences into close proximity.

  • Received:
  • Accepted:
  • Published Online:
© The Journal of General Virology 1994
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-75-10-2645
1994-10-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/75/10/JV0750102645.html?itemId=/content/journal/jgv/10.1099/0022-1317-75-10-2645&mimeType=html&fmt=ahah

References

  1. Astell C. R. 1990; Terminal hairpins OF parvovirus genomes and their role in DNA replication. In Handbook of Parvoviruses 1 pp. 57–79 Tijssen P. Edited by Boca Raton: CRC Press;
     [Google Scholar]
  2. Barrijal S., Perros M., Gu Z., Avalosse B. L., Belenguer P., Amalric F., Rommelaere J. 1992; Nucleolin forms a specific complex with a fragment of the viral (minus) strand of minute virus of mice DNA. Nucleic Acids Research 20:5053–5060
     [Google Scholar]
  3. Ben-Asher E., Aloni Y. 1984; Transcription of minute virus of mice, an autonomous parvovirus, may be regulated by attenuation. Journal of Virology 52:266–276
     [Google Scholar]
  4. Bentley D. L., Groudine M. 1988; Sequence requirements for premature termination of transcription in the human c-myc gene. Cell 53:245–256
     [Google Scholar]
  5. Cheng S.-W. C., Lynch E. C., Leason K. R., Court D. L., Shapirro B. A., Friedman D. I. 1991; Functional importance of sequence in the stem-loop of a transcription terminator. Science 254:1205–1207
     [Google Scholar]
  6. Cornelis J. J., Spruyt N., Spegelaere P., Guetta E., Darawshi T., Cotmore S. F., Tal J., Rommelaere J. 1988; Sensitization of transformed rat fibroblasts to killing by parvovirus minute virus of mice correlates with an increase in viral gene expression. Journal of Virology 62:3438–3444
     [Google Scholar]
  7. Cotmore S. F. 1990; Gene expression in the autonomous parvoviruses. In Handbook of Parvoviruses 1 pp. 141–154 Tijssen P. Edited by Boca Raton: CRC Press;
     [Google Scholar]
  8. Gamper H. B., Hearst J. E. 1982; A topological model for transcription based on unwinding angle analysis of E. coli RNA polymerase binary, initiation and ternary complexes. Cell 29:81–90
     [Google Scholar]
  9. Gluzman Y. 1981; SV40-transformed simian cells support the replication of early SV40 mutants. Cell 23:175–182
     [Google Scholar]
  10. Graham F. L., van der Eb A. J. 1973; A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52:456–457
     [Google Scholar]
  11. Hay N., Aloni Y. 1984; Attenuation in SV40 as a mechanism of transcription-termination by RNA polymerase B. Nucleic Acids Research 12:1401–1414
     [Google Scholar]
  12. Hofstetter H., Schamböck J., van den Berg J., Weissmann C. 1976; Specific excision of the inserted DNA segment from hybrid plasmids constructed by the poly (dA)-poly (dT) method. Biochimica et biophysica acta 454:587–591
     [Google Scholar]
  13. Kessler M., Ben-Asher E., Aloni Y. 1989; Elements modulating the block of transcription elongation at the adenovirus 2 attenuation site. Journal of Biological Chemistry 264:9785–9790
     [Google Scholar]
  14. Krauskopf A., Bengal E., Aloni Y. 1991; The block of transcription elongation at the minute virus of mice attenuator is regulated by cellular elongation factors. Molecular and Cellular Biology 11:3515–3521
     [Google Scholar]
  15. Merchlinsky M. J., Tattersall P. J., Leary J. J., Cotmore S. F., Gardiner E. M., Ward D. C. 1983; Construction of an infectious molecular clone of the autonomous parvovirus minute virus of mice. Journal of Virology 47:227–232
     [Google Scholar]
  16. Moore S. 1981; Pancreatic DNase. In The Enzymes 14 pp. 111–121 Boyer P. D. Edited by San Diego: Academic Press;
     [Google Scholar]
  17. Resnekov O., Aloni Y. 1989; RNA polymerase II is capable of pausing and prematurely terminating transcription at a precise location in vivo and in vitro. Proceedings of the National Academy of Sciences, U.S.A 86:12–16
     [Google Scholar]
  18. Resnekov O., Kessler M., Aloni Y. 1989; RNA secondary structure is an integral part of the in vitro mechanism of attenuation in simian virus 40. Journal of Biological Chemistry 264:9953–9959
     [Google Scholar]
  19. Resnekov O., Pruzan R., Aloni Y. 1991; Elements involved in an in vitro block to transcription elongation at the end of the L1 mRNA family of adenovirus 2. Nucleic Acids Research 19:1783–1790
     [Google Scholar]
  20. Rice G. A., Kane C. M., Chamberlin M. J. 1991; Footprinting analysis of mammalian RNA polymerase II along its transcript: an alternative view of transcription elongation. Proceedings of the National Academy of Sciences, U.S.A 88:4245–4249
     [Google Scholar]
  21. Russell S. J., Brandenburger A., Flemming C. L., Collins M. K. L., Rommelaere J. 1992; Transformation-dependent expression of interleukin genes delivered by a recombinant parvovirus. Journal of Virology 66:2821–2828
     [Google Scholar]
  22. Saluz H., Jost J.-P. 1989; A simple high resolution procedure to study DNA methylation and in vivo DNA-protein interactions on a single-copy gene level in higher eukaryotes. Proceedings of the National Academy of Sciences, U.S.A 86:2602–2606
     [Google Scholar]
  23. Shein H. M., Elders J. F. 1962; Multiplication and cyto-pathogenicity of simian vacuolating virus 40 in cultures of human tissues. Proceedings of the Society of Experimental and Biological Medicine 109:495–500
     [Google Scholar]
  24. Sluder A. E., Price D. H., Greenleaf A. L. 1988; Elongation by Drosophila RNA polymerase II. Journal of Biological Chemistry 263:9917–9925
     [Google Scholar]
  25. Spegelaere P., van Hille B., Spruyt N., Faisst S., Cornelis J. J., Rommelaere J. 1991; Initiation of transcription from the minute virus of mice P4 promoter is stimulated in rat cells expressing a c-Ha-ras oncogene. Journal of Virology 65:4919–4928
     [Google Scholar]
  26. Spencer A., Groudine M. 1990; Transcription elongation and eukaryotic gene regulation. Oncogene 5:777–785
     [Google Scholar]
  27. Suck D., Oefner C. 1986; Structure of DNasel at 2·0 Å resolution suggests a mechanism for binding to and cutting DNA. Nature; London: 321620–625
     [Google Scholar]
  28. Tattersall P., Gardiner E. M. 1990; Autonomous parvovirus-host cell interactions. In Handbook of Parvoviruses 1 pp. 111–121 Tijssen P. Edited by Boca Raton: CRC Press;
     [Google Scholar]
  29. Tijssen P. editor 1990; Introduction: parvoviruses. In Handbook of Parvoviruses 1 pp. 3–7 Raton: CRC Press;
     [Google Scholar]
  30. Toohey M. G., Jones K. A. 1989; In vitro formation of short RNA polymerase II transcripts that terminate within the HIV-1 and HIV-2 promoter-proximal downstream regions. Genes and Development 3:265–282
     [Google Scholar]
  31. van Hille B., Duponchel N., Salome N., Spruyt N., Cotmore S. F., Cornelis J. J., Rommelaere J. 1989; Limitations to the expression of parvoviral nonstructural proteins may determine the extent of sensitization of EJ ras-transformed rat fibroblasts to minute virus of mice. Virology 171:89–97
     [Google Scholar]
  32. Vogt V. 1980; Purification and properties of SI nuclease from Aspergillus. Methods in Enzymology 65:248–255
     [Google Scholar]
  33. Yanofsky C. 1988; Transcription attenuation. Journal of Biological Chemistry 263:609–612
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-75-10-2645
Loading
Cruciform structure of a DNA motif of parvovirus minute virus of mice (prototype strain) involved in the attenuation of gene expression
J. Gen. Virol. 75, 2645 (1994); https://doi.org/10.1099/0022-1317-75-10-2645
/content/journal/jgv/10.1099/0022-1317-75-10-2645
/content/journal/jgv/10.1099/0022-1317-75-10-2645
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error