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A Sub-Element in PRE enhances nuclear export of intronless mRNAs by recruiting the TREX complex via ZC3H18.

Chi B, Wang K, Du Y, Gui B, Chang X, Wang L, Fan J, Chen S, Wu X, Li G, Cheng H - Nucleic Acids Res. (2014)

Bottom Line: We found that PRE drastically enhances the cytoplasmic accumulation of cDNA transcripts independent of any viral protein.Together, our data indicate that SEP1 enhances mRNA export by recruiting TREX via ZC3H18.The new mRNA export factors that we identified might be involved in cap- and splicing-dependent TREX recruitment to cellular mRNAs.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Key Laboratory of Molecular Andrology, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.

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TREX proteins and TAP play critical roles in PRE-mediated mRNA export. (A) HeLa cells were transfected with siRNAs targeting the indicated genes, and lysates were prepared 60 h after transfection. Western analyses of the cell lysates were carried out with the indicated antibodies. Control knockdown cell lysates (10%, 30% and 100%) were loaded. (B) The cG-SEP1 construct was transfected into HeLa cells 36 h after siRNA transfection, and 24 h later, FISH was carried out to detect the cG-SEP1 mRNA. The FISH probe was same as that used in Figure 1. (C) Similar to (B), except that the cG-PRE construct was transfected, and TAP knockdown cells were also included.
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Figure 3: TREX proteins and TAP play critical roles in PRE-mediated mRNA export. (A) HeLa cells were transfected with siRNAs targeting the indicated genes, and lysates were prepared 60 h after transfection. Western analyses of the cell lysates were carried out with the indicated antibodies. Control knockdown cell lysates (10%, 30% and 100%) were loaded. (B) The cG-SEP1 construct was transfected into HeLa cells 36 h after siRNA transfection, and 24 h later, FISH was carried out to detect the cG-SEP1 mRNA. The FISH probe was same as that used in Figure 1. (C) Similar to (B), except that the cG-PRE construct was transfected, and TAP knockdown cells were also included.

Mentions: The fact that TREX specifically associates with the SEP1 RNA raised the possibility that it plays a role in SEP1-dependent mRNA export. To test this possibility, we carried out RNA­-mediated interference (RNAi) of UAP56/URH49, Aly and Thoc2 in HeLa cells and used a non­targeting siRNA as a negative control. Western analyses revealed that protein levels of these genes were efficiently knocked down (Figure 3A). When the cG-SEP1 construct was transfected into control knockdown cells, the corresponding mRNA was mainly detected in the cytoplasm. In marked contrast, the cG-SEP1 mRNA was mostly retained in the nuclei in UAP56/URH49-, Aly- and Thoc2-knockdown cells (Figure 3B). These results indicate that TREX plays key roles in SEP1-mediated mRNA export.


A Sub-Element in PRE enhances nuclear export of intronless mRNAs by recruiting the TREX complex via ZC3H18.

Chi B, Wang K, Du Y, Gui B, Chang X, Wang L, Fan J, Chen S, Wu X, Li G, Cheng H - Nucleic Acids Res. (2014)

TREX proteins and TAP play critical roles in PRE-mediated mRNA export. (A) HeLa cells were transfected with siRNAs targeting the indicated genes, and lysates were prepared 60 h after transfection. Western analyses of the cell lysates were carried out with the indicated antibodies. Control knockdown cell lysates (10%, 30% and 100%) were loaded. (B) The cG-SEP1 construct was transfected into HeLa cells 36 h after siRNA transfection, and 24 h later, FISH was carried out to detect the cG-SEP1 mRNA. The FISH probe was same as that used in Figure 1. (C) Similar to (B), except that the cG-PRE construct was transfected, and TAP knockdown cells were also included.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4066777&req=5

Figure 3: TREX proteins and TAP play critical roles in PRE-mediated mRNA export. (A) HeLa cells were transfected with siRNAs targeting the indicated genes, and lysates were prepared 60 h after transfection. Western analyses of the cell lysates were carried out with the indicated antibodies. Control knockdown cell lysates (10%, 30% and 100%) were loaded. (B) The cG-SEP1 construct was transfected into HeLa cells 36 h after siRNA transfection, and 24 h later, FISH was carried out to detect the cG-SEP1 mRNA. The FISH probe was same as that used in Figure 1. (C) Similar to (B), except that the cG-PRE construct was transfected, and TAP knockdown cells were also included.
Mentions: The fact that TREX specifically associates with the SEP1 RNA raised the possibility that it plays a role in SEP1-dependent mRNA export. To test this possibility, we carried out RNA­-mediated interference (RNAi) of UAP56/URH49, Aly and Thoc2 in HeLa cells and used a non­targeting siRNA as a negative control. Western analyses revealed that protein levels of these genes were efficiently knocked down (Figure 3A). When the cG-SEP1 construct was transfected into control knockdown cells, the corresponding mRNA was mainly detected in the cytoplasm. In marked contrast, the cG-SEP1 mRNA was mostly retained in the nuclei in UAP56/URH49-, Aly- and Thoc2-knockdown cells (Figure 3B). These results indicate that TREX plays key roles in SEP1-mediated mRNA export.

Bottom Line: We found that PRE drastically enhances the cytoplasmic accumulation of cDNA transcripts independent of any viral protein.Together, our data indicate that SEP1 enhances mRNA export by recruiting TREX via ZC3H18.The new mRNA export factors that we identified might be involved in cap- and splicing-dependent TREX recruitment to cellular mRNAs.

View Article: PubMed Central - PubMed

Affiliation: Shanghai Key Laboratory of Molecular Andrology, State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.

Show MeSH
Related in: MedlinePlus