PMID- 19365637
OWN - NLM
STAT- MEDLINE
DCOM- 20090831
LR  - 20240213
IS  - 1432-119X (Electronic)
IS  - 0948-6143 (Print)
IS  - 0948-6143 (Linking)
VI  - 132
IP  - 1
DP  - 2009 Jul
TI  - LPA-induced mutually exclusive subcellular localization of active RhoA and Arp2 
      mRNA revealed by sequential FRET and FISH.
PG  - 47-58
LID - 10.1007/s00418-009-0589-x [doi]
AB  - We previously demonstrated that mRNAs for the subunits of the Arp2/3 complex 
      localize to protrusions in fibroblasts (Mingle et al. in J Cell Sci 
      118:2425-2433, 2005). However, the signaling pathway that regulates Arp2/3 
      complex mRNA localization remains unknown. In this study we have identified 
      lysophosphatidic acid (LPA) as a potent inducer of Arp2 mRNA localization to 
      protrusions in fibroblasts via the RhoA-ROCK pathway. As RhoA is known to be 
      activated locally in the cells, we sought to understand how spatial activation of 
      Rho affects Arp2 mRNA localization. By sequentially performing fluorescence 
      resonance energy transfer (FRET) and fluorescence in situ hybridization (FISH), 
      we have visualized active RhoA and Arp2 mRNA in the same cells. Upon LPA 
      stimulation, approximately two times more cells than those in the serum-free 
      medium showed mutually exclusive localization of active RhoA and Arp2 mRNA. These 
      results demonstrate the importance of localized activation of Rho in Arp2 mRNA 
      localization and provide new insights as to how Rho regulates Arp2/3 complex mRNA 
      localization. To our best knowledge, this is the first report in which FRET and 
      FISH are combined to detect localized protein activity and mRNA in the same 
      cells. This method should be easily adopted for the detection of other 
      fluorescence protein based biosensors and DNA/RNA in the same cells.
FAU - Mingle, Lisa A
AU  - Mingle LA
AD  - Center for Cell Biology and Cancer Research, Albany Medical College, 47 New 
      Scotland Avenue, Albany, NY 12208, USA.
FAU - Bonamy, Ghislain
AU  - Bonamy G
FAU - Barroso, Margarida
AU  - Barroso M
FAU - Liao, Guoning
AU  - Liao G
FAU - Liu, Gang
AU  - Liu G
LA  - eng
GR  - R01 GM070560-04/GM/NIGMS NIH HHS/United States
GR  - T32 HL007194/HL/NHLBI NIH HHS/United States
GR  - T32-HL-07194/HL/NHLBI NIH HHS/United States
GR  - R01 GM070560/GM/NIGMS NIH HHS/United States
GR  - GM70560/GM/NIGMS NIH HHS/United States
GR  - R01 GM070560-05/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20090414
PL  - Germany
TA  - Histochem Cell Biol
JT  - Histochemistry and cell biology
JID - 9506663
RN  - 0 (Actin-Related Protein 2)
RN  - 0 (Lysophospholipids)
RN  - 0 (RNA, Messenger)
RN  - EC 3.6.5.2 (rhoA GTP-Binding Protein)
RN  - PG6M3969SG (lysophosphatidic acid)
SB  - IM
MH  - Actin-Related Protein 2/*metabolism
MH  - Animals
MH  - Cell Nucleus/metabolism
MH  - Cells, Cultured
MH  - Chick Embryo
MH  - Cytoplasm/metabolism
MH  - Fibroblasts/cytology/drug effects/*metabolism
MH  - Fluorescence Resonance Energy Transfer
MH  - In Situ Hybridization, Fluorescence
MH  - Lysophospholipids/*pharmacology
MH  - RNA, Messenger/*metabolism
MH  - rhoA GTP-Binding Protein/*metabolism
PMC - PMC2753266
MID - NIHMS141990
EDAT- 2009/04/15 09:00
MHDA- 2009/09/01 06:00
PMCR- 2010/01/01
CRDT- 2009/04/15 09:00
PHST- 2009/03/18 00:00 [accepted]
PHST- 2009/04/15 09:00 [entrez]
PHST- 2009/04/15 09:00 [pubmed]
PHST- 2009/09/01 06:00 [medline]
PHST- 2010/01/01 00:00 [pmc-release]
AID - 10.1007/s00418-009-0589-x [doi]
PST - ppublish
SO  - Histochem Cell Biol. 2009 Jul;132(1):47-58. doi: 10.1007/s00418-009-0589-x. Epub 
      2009 Apr 14.