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The closely related RNA helicases, UAP56 and URH49, preferentially form distinct mRNA export machineries and coordinately regulate mitotic progression.

Yamazaki T, Fujiwara N, Yukinaga H, Ebisuya M, Shiki T, Kurihara T, Kioka N, Kambe T, Nagao M, Nishida E, Masuda S - Mol. Biol. Cell (2010)

Bottom Line: Consistent with their target mRNAs, depletion of UAP56 causes mitotic delay and sister chromatid cohesion defects, whereas depletion of URH49 causes chromosome arm resolution defects and failure of cytokinesis.In addition, depletion of the other human TREX components or CIP29 causes mitotic defects similar to those observed in UAP56- or URH49-depleted cells, respectively.Taken together, the two closely related RNA helicases have evolved to form distinct mRNA export machineries, which regulate mitosis at different steps.

View Article: PubMed Central - PubMed

Affiliation: Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kyoto, Japan.

ABSTRACT
Nuclear export of mRNA is an essential process for eukaryotic gene expression. The TREX complex couples gene expression from transcription and splicing to mRNA export. Sub2, a core component of the TREX complex in yeast, has diversified in humans to two closely related RNA helicases, UAP56 and URH49. Here, we show that URH49 forms a novel URH49-CIP29 complex, termed the AREX (alternative mRNA export) complex, whereas UAP56 forms the human TREX complex. The mRNAs regulated by these helicases are different at the genome-wide level. The two sets of target mRNAs contain distinct subsets of key mitotic regulators. Consistent with their target mRNAs, depletion of UAP56 causes mitotic delay and sister chromatid cohesion defects, whereas depletion of URH49 causes chromosome arm resolution defects and failure of cytokinesis. In addition, depletion of the other human TREX components or CIP29 causes mitotic defects similar to those observed in UAP56- or URH49-depleted cells, respectively. Taken together, the two closely related RNA helicases have evolved to form distinct mRNA export machineries, which regulate mitosis at different steps.

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UAP56 forms the hTREX complex, whereas URH49 forms the AREX complex. (A) Top, UAP56 and URH49 both localize to the nucleus. HeLa cells were stained with anti-UAP56- or anti-URH49-specific antibody. Bottom, EGFP-UAP56 and DsRed-URH49 colocalized in the nucleus. Scale bar, 10 μm. (B) UAP56 interacted with the hTHO complex and Aly, whereas URH49 bound to CIP29 in vivo. Endogenous UAP56 and URH49 were immunoprecipitated with specific antibodies, and the immunoprecipitates were analyzed by immunoblotting with the indicated antibodies. Control 1 indicates preimmune serum from rat. IP, immunoprecipitation; IB, immunoblotting. (C) The hTHO complex and Aly coimmunoprecipitated the hTREX components, whereas CIP29 coimmunoprecipitated URH49. Endogenous hTho2, fSAP79, Aly, and CIP29 were immunoprecipitated with specific antibodies, and the immunoprecipitates were analyzed by immunoblotting with the indicated antibodies. Controls 1 and 2 indicate pre immune serum from rat and rabbit, respectively. IP, immunoprecipitation; IB, immunoblotting. (D) Schematic representation of the hTREX complex and the AREX complex.
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Figure 5: UAP56 forms the hTREX complex, whereas URH49 forms the AREX complex. (A) Top, UAP56 and URH49 both localize to the nucleus. HeLa cells were stained with anti-UAP56- or anti-URH49-specific antibody. Bottom, EGFP-UAP56 and DsRed-URH49 colocalized in the nucleus. Scale bar, 10 μm. (B) UAP56 interacted with the hTHO complex and Aly, whereas URH49 bound to CIP29 in vivo. Endogenous UAP56 and URH49 were immunoprecipitated with specific antibodies, and the immunoprecipitates were analyzed by immunoblotting with the indicated antibodies. Control 1 indicates preimmune serum from rat. IP, immunoprecipitation; IB, immunoblotting. (C) The hTHO complex and Aly coimmunoprecipitated the hTREX components, whereas CIP29 coimmunoprecipitated URH49. Endogenous hTho2, fSAP79, Aly, and CIP29 were immunoprecipitated with specific antibodies, and the immunoprecipitates were analyzed by immunoblotting with the indicated antibodies. Controls 1 and 2 indicate pre immune serum from rat and rabbit, respectively. IP, immunoprecipitation; IB, immunoblotting. (D) Schematic representation of the hTREX complex and the AREX complex.

Mentions: To investigate the differences between UAP56 and URH49, their cellular localization was examined. Endogenous (Figure 5A) and FLAG-tagged UAP56 and URH49 (Supplemental Figure S4A) localized to the nucleus, as previously described (Kota et al., 2008). EGFP-UAP56 and DsRed-URH49 also colocalized to the nucleus (Figure 5A). These data indicate that UAP56 and URH49 localized in the same, nuclear compartment.


The closely related RNA helicases, UAP56 and URH49, preferentially form distinct mRNA export machineries and coordinately regulate mitotic progression.

Yamazaki T, Fujiwara N, Yukinaga H, Ebisuya M, Shiki T, Kurihara T, Kioka N, Kambe T, Nagao M, Nishida E, Masuda S - Mol. Biol. Cell (2010)

UAP56 forms the hTREX complex, whereas URH49 forms the AREX complex. (A) Top, UAP56 and URH49 both localize to the nucleus. HeLa cells were stained with anti-UAP56- or anti-URH49-specific antibody. Bottom, EGFP-UAP56 and DsRed-URH49 colocalized in the nucleus. Scale bar, 10 μm. (B) UAP56 interacted with the hTHO complex and Aly, whereas URH49 bound to CIP29 in vivo. Endogenous UAP56 and URH49 were immunoprecipitated with specific antibodies, and the immunoprecipitates were analyzed by immunoblotting with the indicated antibodies. Control 1 indicates preimmune serum from rat. IP, immunoprecipitation; IB, immunoblotting. (C) The hTHO complex and Aly coimmunoprecipitated the hTREX components, whereas CIP29 coimmunoprecipitated URH49. Endogenous hTho2, fSAP79, Aly, and CIP29 were immunoprecipitated with specific antibodies, and the immunoprecipitates were analyzed by immunoblotting with the indicated antibodies. Controls 1 and 2 indicate pre immune serum from rat and rabbit, respectively. IP, immunoprecipitation; IB, immunoblotting. (D) Schematic representation of the hTREX complex and the AREX complex.
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Figure 5: UAP56 forms the hTREX complex, whereas URH49 forms the AREX complex. (A) Top, UAP56 and URH49 both localize to the nucleus. HeLa cells were stained with anti-UAP56- or anti-URH49-specific antibody. Bottom, EGFP-UAP56 and DsRed-URH49 colocalized in the nucleus. Scale bar, 10 μm. (B) UAP56 interacted with the hTHO complex and Aly, whereas URH49 bound to CIP29 in vivo. Endogenous UAP56 and URH49 were immunoprecipitated with specific antibodies, and the immunoprecipitates were analyzed by immunoblotting with the indicated antibodies. Control 1 indicates preimmune serum from rat. IP, immunoprecipitation; IB, immunoblotting. (C) The hTHO complex and Aly coimmunoprecipitated the hTREX components, whereas CIP29 coimmunoprecipitated URH49. Endogenous hTho2, fSAP79, Aly, and CIP29 were immunoprecipitated with specific antibodies, and the immunoprecipitates were analyzed by immunoblotting with the indicated antibodies. Controls 1 and 2 indicate pre immune serum from rat and rabbit, respectively. IP, immunoprecipitation; IB, immunoblotting. (D) Schematic representation of the hTREX complex and the AREX complex.
Mentions: To investigate the differences between UAP56 and URH49, their cellular localization was examined. Endogenous (Figure 5A) and FLAG-tagged UAP56 and URH49 (Supplemental Figure S4A) localized to the nucleus, as previously described (Kota et al., 2008). EGFP-UAP56 and DsRed-URH49 also colocalized to the nucleus (Figure 5A). These data indicate that UAP56 and URH49 localized in the same, nuclear compartment.

Bottom Line: Consistent with their target mRNAs, depletion of UAP56 causes mitotic delay and sister chromatid cohesion defects, whereas depletion of URH49 causes chromosome arm resolution defects and failure of cytokinesis.In addition, depletion of the other human TREX components or CIP29 causes mitotic defects similar to those observed in UAP56- or URH49-depleted cells, respectively.Taken together, the two closely related RNA helicases have evolved to form distinct mRNA export machineries, which regulate mitosis at different steps.

View Article: PubMed Central - PubMed

Affiliation: Division of Integrated Life Science, Graduate School of Biostudies, Kyoto University, Kyoto, Japan.

ABSTRACT
Nuclear export of mRNA is an essential process for eukaryotic gene expression. The TREX complex couples gene expression from transcription and splicing to mRNA export. Sub2, a core component of the TREX complex in yeast, has diversified in humans to two closely related RNA helicases, UAP56 and URH49. Here, we show that URH49 forms a novel URH49-CIP29 complex, termed the AREX (alternative mRNA export) complex, whereas UAP56 forms the human TREX complex. The mRNAs regulated by these helicases are different at the genome-wide level. The two sets of target mRNAs contain distinct subsets of key mitotic regulators. Consistent with their target mRNAs, depletion of UAP56 causes mitotic delay and sister chromatid cohesion defects, whereas depletion of URH49 causes chromosome arm resolution defects and failure of cytokinesis. In addition, depletion of the other human TREX components or CIP29 causes mitotic defects similar to those observed in UAP56- or URH49-depleted cells, respectively. Taken together, the two closely related RNA helicases have evolved to form distinct mRNA export machineries, which regulate mitosis at different steps.

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