Stable Assembly of HIV-1 Export Complexes Occurs Cotranscriptionally

Overview

Journal RNA

Specialty Molecular Biology

Date 2013 Nov 21

PMID 24255166

Citations 16

Authors

Isabel Nawroth

Florian Mueller

Eugenia Basyuk

Nancy Beerens

Ulrik L Rahbek

Xavier Darzacq

Edouard Bertrand

Jorgen Kjems

Ute Schmidt

Affiliations

Soon will be listed here.

Abstract

The HIV-1 Rev protein mediates export of unspliced and singly spliced viral transcripts by binding to the Rev response element (RRE) and recruiting the cellular export factor CRM1. Here, we investigated the recruitment of Rev to the transcription sites of HIV-1 reporters that splice either post- or cotranscriptionally. In both cases, we observed that Rev localized to the transcription sites of the reporters and recruited CRM1. Rev and CRM1 remained at the reporter transcription sites when cells were treated with the splicing inhibitor Spliceostatin A (SSA), showing that the proteins associate with RNA prior to or during early spliceosome assembly. Fluorescence recovery after photobleaching (FRAP) revealed that Rev and CRM1 have similar kinetics as the HIV-1 RNA, indicating that Rev, CRM1, and RRE-containing RNAs are released from the site of transcription in one single export complex. These results suggest that cotranscriptional formation of a stable export complex serves as a means to ensure efficient export of unspliced viral RNAs.

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References

Sherer N, Swanson C, Hue S, Roberts R, Bergeron J, Malim M . Evolution of a species-specific determinant within human CRM1 that regulates the post-transcriptional phases of HIV-1 replication. PLoS Pathog. 2011; 7(11):e1002395. PMC: 3219727. DOI: 10.1371/journal.ppat.1002395. View

Chang D, Sharp P . Regulation by HIV Rev depends upon recognition of splice sites. Cell. 1989; 59(5):789-95. DOI: 10.1016/0092-8674(89)90602-8. View

Hammarskjold M, Li H, Rekosh D, Prasad S . Human immunodeficiency virus env expression becomes Rev-independent if the env region is not defined as an intron. J Virol. 1994; 68(2):951-8. PMC: 236533. DOI: 10.1128/JVI.68.2.951-958.1994. View

Yi R, Bogerd H, Cullen B . Recruitment of the Crm1 nuclear export factor is sufficient to induce cytoplasmic expression of incompletely spliced human immunodeficiency virus mRNAs. J Virol. 2002; 76(5):2036-42. PMC: 153812. DOI: 10.1128/jvi.76.5.2036-2042.2002. View

Molle D, Maiuri P, Boireau S, Bertrand E, Knezevich A, Marcello A . A real-time view of the TAR:Tat:P-TEFb complex at HIV-1 transcription sites. Retrovirology. 2007; 4:36. PMC: 1904240. DOI: 10.1186/1742-4690-4-36. View

Kammler S, Otte M, Hauber I, Kjems J, Hauber J, Schaal H . The strength of the HIV-1 3' splice sites affects Rev function. Retrovirology. 2006; 3:89. PMC: 1697824. DOI: 10.1186/1742-4690-3-89. View

Kaida D, Motoyoshi H, Tashiro E, Nojima T, Hagiwara M, Ishigami K . Spliceostatin A targets SF3b and inhibits both splicing and nuclear retention of pre-mRNA. Nat Chem Biol. 2007; 3(9):576-83. DOI: 10.1038/nchembio.2007.18. View

Bertrand E, Chartrand P, Schaefer M, Shenoy S, Singer R, Long R . Localization of ASH1 mRNA particles in living yeast. Mol Cell. 1998; 2(4):437-45. DOI: 10.1016/s1097-2765(00)80143-4. View

Kjems J, Sharp P . The basic domain of Rev from human immunodeficiency virus type 1 specifically blocks the entry of U4/U6.U5 small nuclear ribonucleoprotein in spliceosome assembly. J Virol. 1993; 67(8):4769-76. PMC: 237863. DOI: 10.1128/JVI.67.8.4769-4776.1993. View

10.

Kjems J, Frankel A, Sharp P . Specific regulation of mRNA splicing in vitro by a peptide from HIV-1 Rev. Cell. 1991; 67(1):169-78. DOI: 10.1016/0092-8674(91)90580-r. View

11.

Kjems J . Inefficient spliceosome assembly and abnormal branch site selection in splicing of an HIV-1 transcript in vitro. J Biol Chem. 1995; 270(41):24060-6. DOI: 10.1074/jbc.270.41.24060. View

12.

Roybal G, Jurica M . Spliceostatin A inhibits spliceosome assembly subsequent to prespliceosome formation. Nucleic Acids Res. 2010; 38(19):6664-72. PMC: 2965229. DOI: 10.1093/nar/gkq494. View

13.

Daelemans D, Costes S, Lockett S, Pavlakis G . Kinetic and molecular analysis of nuclear export factor CRM1 association with its cargo in vivo. Mol Cell Biol. 2005; 25(2):728-39. PMC: 543413. DOI: 10.1128/MCB.25.2.728-739.2005. View

14.

Stoltzfus C, Madsen J . Role of viral splicing elements and cellular RNA binding proteins in regulation of HIV-1 alternative RNA splicing. Curr HIV Res. 2006; 4(1):43-55. DOI: 10.2174/157016206775197655. View

15.

Mueller F, Mazza D, Stasevich T, McNally J . FRAP and kinetic modeling in the analysis of nuclear protein dynamics: what do we really know?. Curr Opin Cell Biol. 2010; 22(3):403-11. PMC: 2916960. DOI: 10.1016/j.ceb.2010.03.002. View

16.

Schmidt U, Basyuk E, Robert M, Yoshida M, Villemin J, Auboeuf D . Real-time imaging of cotranscriptional splicing reveals a kinetic model that reduces noise: implications for alternative splicing regulation. J Cell Biol. 2011; 193(5):819-29. PMC: 3105549. DOI: 10.1083/jcb.201009012. View

17.

Zillmann M, Zapp M, Berget S . Gel electrophoretic isolation of splicing complexes containing U1 small nuclear ribonucleoprotein particles. Mol Cell Biol. 1988; 8(2):814-21. PMC: 363208. DOI: 10.1128/mcb.8.2.814-821.1988. View

18.

Neville M, Stutz F, Lee L, Davis L, Rosbash M . The importin-beta family member Crm1p bridges the interaction between Rev and the nuclear pore complex during nuclear export. Curr Biol. 1997; 7(10):767-75. DOI: 10.1016/s0960-9822(06)00335-6. View

19.

Fornerod M, Ohno M, Yoshida M, Mattaj I . CRM1 is an export receptor for leucine-rich nuclear export signals. Cell. 1997; 90(6):1051-60. DOI: 10.1016/s0092-8674(00)80371-2. View

20.

Boireau S, Maiuri P, Basyuk E, Mata M, Knezevich A, Pradet-Balade B . The transcriptional cycle of HIV-1 in real-time and live cells. J Cell Biol. 2007; 179(2):291-304. PMC: 2064765. DOI: 10.1083/jcb.200706018. View