A role for nucleoporin FG repeat domains in export of human immunodeficiency virus type 1 Rev protein and RNA from the nucleus

The human immunodeficiency virus type 1 Rev protein contains a nuclear export signal (NES) that is required for Rev-mediated RNA export in mammals as well as in the yeast Saccharomyces cerevisiae. The Rev NES has been shown to specifically interact with a human (hRIP/RAB1) and a yeast (yRip1p) protein in the two-hybrid assay. Both of these interacting proteins are related to FG nucleoporins on the basis of the presence of typical repeat motifs. This paper shows that Rev is able to interact with multiple FG repeat-containing nucleoporins from both S. cerevisiae and mammals; moreover, the ability of Rev NES mutants to interact with these FG nucleoporins parallels the ability of the mutants to promote RNA export in yeast and mammalian cells. The data also show that, after Xenopus oocyte nuclear injection, several FG nucleoporin repeat domains inhibit the export of both Rev protein and U small nuclear RNAs, suggesting that these nucleoporins participate in Rev-mediated and cellular RNA export. Interestingly, not all FG nucleoporin repeat domains produced the same pattern of RNA export inhibition. The results suggest that Rev and cellular mediators of RNA export can interact with multiple components of the nuclear pore complex during transport, analogous to the proposed mode of action of the nuclear protein import receptor.

[1]  Utz Fischer,et al.  The HIV-1 Rev Activation Domain is a nuclear export signal that accesses an export pathway used by specific cellular RNAs , 1995, Cell.

[2]  G. Blobel,et al.  The human CAN protein, a putative oncogene product associated with myeloid leukemogenesis, is a nuclear pore complex protein that faces the cytoplasm. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[3]  G. Blobel,et al.  The Essential Yeast Nucleoporin NUP159 Is Located on the Cytoplasmic Side of the Nuclear Pore Complex and Serves in Karyopherin-mediated Binding of Transport Substrate (*) , 1995, The Journal of Biological Chemistry.

[4]  R. Brent,et al.  Interaction mating reveals binary and ternary connections between Drosophila cell cycle regulators. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[5]  C. Kambach,et al.  Intracellular distribution of the U1A protein depends on active transport and nuclear binding to U1 snRNA , 1992, The Journal of cell biology.

[6]  P. Legrain,et al.  Some cis- and trans-acting mutants for splicing target pre-mRNA to the cytoplasm , 1989, Cell.

[7]  Michael R. Green,et al.  A human nucleoporin-like protein that specifically interacts with HIV Rev , 1995, Nature.

[8]  Joe D. Lewis,et al.  A cap-binding protein complex mediating U snRNA export , 1995, Nature.

[9]  M. Malim,et al.  Amphibian transcription factor IIIA proteins contain a sequence element functionally equivalent to the nuclear export signal of human immunodeficiency virus type 1 Rev. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[10]  D. Smith,et al.  Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. , 1988, Gene.

[11]  Roger Brent,et al.  C dil, a Human Gl and S Phase Protein Phosphatase That Associates with Cdk2 , 2003 .

[12]  M. Rosenberg,et al.  Rev-dependent expression of human immunodeficiency virus type 1 gp160 in Drosophila melanogaster cells , 1990, Molecular and cellular biology.

[13]  G. Blobel,et al.  Protein import into nuclei: association and dissociation reactions involving transport substrate, transport factors, and nucleoporins , 1995, Cell.

[14]  C. Cole,et al.  A conditional allele of the novel repeat-containing yeast nucleoporin RAT7/NUP159 causes both rapid cessation of mRNA export and reversible clustering of nuclear pore complexes , 1995, The Journal of cell biology.

[15]  G. Fink,et al.  NUP2, a novel yeast nucleoporin, has functional overlap with other proteins of the nuclear pore complex. , 1993, Molecular biology of the cell.

[16]  W. Boelens,et al.  Nuclear export of different classes of RNA is mediated by specific factors , 1994, The Journal of cell biology.

[17]  G. Fink,et al.  The NUP1 gene encodes an essential component of the yeast nuclear pore complex , 1990, Cell.

[18]  Michael R. Green,et al.  HIV Rev uses a conserved cellular protein export pathway for the nucleocytoplasmic transport of viral RNAs , 1996, Current Biology.

[19]  G. Blobel,et al.  A new family of yeast nuclear pore complex proteins , 1992, The Journal of cell biology.

[20]  G Rautmann,et al.  Evidence that HIV‐1 Rev directly promotes the nuclear export of unspliced RNA. , 1994, The EMBO journal.

[21]  G. Blobel,et al.  Identification of a protein complex that is required for nuclear protein import and mediates docking of import substrate to distinct nucleoporins. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[22]  M. Rout,et al.  Pores for thought: nuclear pore complex proteins. , 1994, Trends in cell biology.

[23]  J. Hauber,et al.  Inhibition of HIV-1 Replication in Lymphocytes by Mutants of the Rev Cofactor eIF-5A , 1996, Science.

[24]  E. Hurt,et al.  Nup145p is required for nuclear export of mRNA and binds homopolymeric RNA in vitro via a novel conserved motif , 1994, Cell.

[25]  G. Blobel,et al.  NUP145 encodes a novel yeast glycine-leucine-phenylalanine-glycine (GLFG) nucleoporin required for nuclear envelope structure , 1994, The Journal of cell biology.

[26]  P. Grandi,et al.  A new subclass of nucleoporins that functionally interact with nuclear pore protein NSP1. , 1992, The EMBO journal.

[27]  M. Malim,et al.  The HIV-1 Rev protein: prototype of a novel class of eukaryotic post-transcriptional regulators. , 1991, Trends in biochemical sciences.

[28]  M. Rosbash,et al.  Identification of a novel nuclear pore-associated protein as a functional target of the HIV-1 Rev protein in yeast , 1995, Cell.

[29]  J. Hauber,et al.  Eukaryotic initiation factor 5A is a cellular target of the human immunodeficiency virus type 1 Rev activation domain mediating trans- activation , 1993, The Journal of cell biology.

[30]  G. Blobel,et al.  The peptide repeat domain of nucleoporin Nup98 functions as a docking site in transport across the nuclear pore complex , 1995, Cell.

[31]  B. Cullen,et al.  Nuclear export of late HIV-1 mRNAs occurs via a cellular protein export pathway. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[32]  K. Murata,et al.  Transformation of intact yeast cells treated with alkali cations. , 1984, Journal of bacteriology.

[33]  E. Hurt,et al.  A novel nucleoskeletal‐like protein located at the nuclear periphery is required for the life cycle of Saccharomyces cerevisiae. , 1988, The EMBO journal.

[34]  P. Grandi,et al.  Functional interaction of Nic96p with a core nucleoporin complex consisting of Nsp1p, Nup49p and a novel protein Nup57p. , 1995, The EMBO journal.

[35]  Roger Y Tsien,et al.  Identification of a signal for rapid export of proteins from the nucleus , 1995, Cell.

[36]  M. Malim,et al.  Mutational definition of the human immunodeficiency virus type 1 Rev activation domain , 1991, Journal of virology.

[37]  M. Rosbash,et al.  A functional interaction between Rev and yeast pre‐mRNA is related to splicing complex formation. , 1994, The EMBO journal.

[38]  Deutsches Krebsforschungszentrum,et al.  Nucleocytoplasmic Transport , 1986, Springer Berlin Heidelberg.

[39]  J. L. Watkins,et al.  The GLFG repetitive region of the nucleoporin Nup116p interacts with Kap95p, an essential yeast nuclear import factor , 1995, The Journal of cell biology.

[40]  B. Cullen,et al.  Identification of a novel cellular cofactor for the Rev/Rex class of retroviral regulatory proteins , 1995, Cell.