Limits...
Inter-cellular transport of ran GTPase.

Khuperkar D, Helen M, Magre I, Joseph J - PLoS ONE (2015)

Bottom Line: Over-expression of Ran is associated with various cancers, although the molecular mechanism underlying this phenomenon is unclear.Serendipitously, we found that Ran possesses the ability to move from cell-to-cell when transiently expressed in mammalian cells.Interestingly, leptomycin B, an inhibitor of CRM1-mediated export, or siRNA mediated depletion of CRM1, significantly impaired the inter-cellular transport of Ran, suggesting a function for CRM1 in this process.

View Article: PubMed Central - PubMed

Affiliation: National Centre for Cell Science, Ganeshkhind, Pune, India.

ABSTRACT
Ran, a member of the Ras-GTPase superfamily, has a well-established role in regulating the transport of macromolecules across the nuclear envelope (NE). Ran has also been implicated in mitosis, cell cycle progression, and NE formation. Over-expression of Ran is associated with various cancers, although the molecular mechanism underlying this phenomenon is unclear. Serendipitously, we found that Ran possesses the ability to move from cell-to-cell when transiently expressed in mammalian cells. Moreover, we show that the inter-cellular transport of Ran is GTP-dependent. Importantly, Ran displays a similar distribution pattern in the recipient cells as that in the donor cell and co-localizes with the Ran binding protein Nup358 (also called RanBP2). Interestingly, leptomycin B, an inhibitor of CRM1-mediated export, or siRNA mediated depletion of CRM1, significantly impaired the inter-cellular transport of Ran, suggesting a function for CRM1 in this process. These novel findings indicate a possible role for Ran beyond nucleo-cytoplasmic transport, with potential implications in inter-cellular communication and cancers.

No MeSH data available.


Related in: MedlinePlus

RanQ69L co-localizes with Nup358 in the recipient cells.HeLa cells were transfected with HA-GAPDH (transfection marker) and 2xGFP-control or Ran mutants as indicated. Cells were fixed and stained for HA-GAPDH (blue) and endogenous Nup358 (Endo.Nup358, red) using specific antibodies. GFP (green) was visualized by direct epifluorescence. Arrow indicates co-localization of RanQ69L with Nup358 on nuclear envelope and arrow head indicates co-localization in cytoplasmic punctae of recipient cells. Scale bar, 20 μm.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4403925&req=5

pone.0125506.g004: RanQ69L co-localizes with Nup358 in the recipient cells.HeLa cells were transfected with HA-GAPDH (transfection marker) and 2xGFP-control or Ran mutants as indicated. Cells were fixed and stained for HA-GAPDH (blue) and endogenous Nup358 (Endo.Nup358, red) using specific antibodies. GFP (green) was visualized by direct epifluorescence. Arrow indicates co-localization of RanQ69L with Nup358 on nuclear envelope and arrow head indicates co-localization in cytoplasmic punctae of recipient cells. Scale bar, 20 μm.

Mentions: Nup358 has been identified as a major Ran-GTP binding protein in the cytoplasm [46,47]. A closer look at the distribution of Ran-Q69L revealed that it is predominantly present on the NE and in cytoplasmic punctae of the acceptor cell (Fig 4). Co-localization studies clearly indicated that Ran-Q69L is present along with Nup358 both on the NE and in the cytoplasmic punctae of the acceptor cells (Fig 4, arrow and arrowheads, respectively). GFP-control and GFP-Ran-T24N, however, did not show any significant co-localization with Nup358 in the cytoplasm. These data suggest that Ran-Q69L present in the recipient cell functionally retains the ability to bind to its partners such as Nup358.


Inter-cellular transport of ran GTPase.

Khuperkar D, Helen M, Magre I, Joseph J - PLoS ONE (2015)

RanQ69L co-localizes with Nup358 in the recipient cells.HeLa cells were transfected with HA-GAPDH (transfection marker) and 2xGFP-control or Ran mutants as indicated. Cells were fixed and stained for HA-GAPDH (blue) and endogenous Nup358 (Endo.Nup358, red) using specific antibodies. GFP (green) was visualized by direct epifluorescence. Arrow indicates co-localization of RanQ69L with Nup358 on nuclear envelope and arrow head indicates co-localization in cytoplasmic punctae of recipient cells. Scale bar, 20 μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4403925&req=5

pone.0125506.g004: RanQ69L co-localizes with Nup358 in the recipient cells.HeLa cells were transfected with HA-GAPDH (transfection marker) and 2xGFP-control or Ran mutants as indicated. Cells were fixed and stained for HA-GAPDH (blue) and endogenous Nup358 (Endo.Nup358, red) using specific antibodies. GFP (green) was visualized by direct epifluorescence. Arrow indicates co-localization of RanQ69L with Nup358 on nuclear envelope and arrow head indicates co-localization in cytoplasmic punctae of recipient cells. Scale bar, 20 μm.
Mentions: Nup358 has been identified as a major Ran-GTP binding protein in the cytoplasm [46,47]. A closer look at the distribution of Ran-Q69L revealed that it is predominantly present on the NE and in cytoplasmic punctae of the acceptor cell (Fig 4). Co-localization studies clearly indicated that Ran-Q69L is present along with Nup358 both on the NE and in the cytoplasmic punctae of the acceptor cells (Fig 4, arrow and arrowheads, respectively). GFP-control and GFP-Ran-T24N, however, did not show any significant co-localization with Nup358 in the cytoplasm. These data suggest that Ran-Q69L present in the recipient cell functionally retains the ability to bind to its partners such as Nup358.

Bottom Line: Over-expression of Ran is associated with various cancers, although the molecular mechanism underlying this phenomenon is unclear.Serendipitously, we found that Ran possesses the ability to move from cell-to-cell when transiently expressed in mammalian cells.Interestingly, leptomycin B, an inhibitor of CRM1-mediated export, or siRNA mediated depletion of CRM1, significantly impaired the inter-cellular transport of Ran, suggesting a function for CRM1 in this process.

View Article: PubMed Central - PubMed

Affiliation: National Centre for Cell Science, Ganeshkhind, Pune, India.

ABSTRACT
Ran, a member of the Ras-GTPase superfamily, has a well-established role in regulating the transport of macromolecules across the nuclear envelope (NE). Ran has also been implicated in mitosis, cell cycle progression, and NE formation. Over-expression of Ran is associated with various cancers, although the molecular mechanism underlying this phenomenon is unclear. Serendipitously, we found that Ran possesses the ability to move from cell-to-cell when transiently expressed in mammalian cells. Moreover, we show that the inter-cellular transport of Ran is GTP-dependent. Importantly, Ran displays a similar distribution pattern in the recipient cells as that in the donor cell and co-localizes with the Ran binding protein Nup358 (also called RanBP2). Interestingly, leptomycin B, an inhibitor of CRM1-mediated export, or siRNA mediated depletion of CRM1, significantly impaired the inter-cellular transport of Ran, suggesting a function for CRM1 in this process. These novel findings indicate a possible role for Ran beyond nucleo-cytoplasmic transport, with potential implications in inter-cellular communication and cancers.

No MeSH data available.


Related in: MedlinePlus