PMID- 10614948
OWN - NLM
STAT- MEDLINE
DCOM- 20000215
LR  - 20190815
IS  - 0142-9612 (Print)
IS  - 0142-9612 (Linking)
VI  - 20
IP  - 23-24
DP  - 1999 Dec
TI  - RhoA-induced changes in fibroblasts cultured on organic monolayers.
PG  - 2435-46
AB  - Substantial previous work indicates that adherent cell morphology in culture is 
      modulated by surface chemistry. Activation of the intracellular small molecular 
      weight GTPase, RhoA, has recently been shown to play an essential role in 
      controlling initiation of key integrin-mediated events in surface adhesion and 
      proliferation. RhoA is interconvertible between an active, membrane-bound form 
      and an inactive, cytosolic RhoGDI-bound form in response to integrin stimulation. 
      This study reports the use of self-assembled functionalized organic alkylthiol 
      monolayers (SAMs) as well-defined cell culture substrates to investigate the 
      relationships between surface chemistry, RhoA activation and subsequent cell 
      morphological and molecular level signal transduction responses in cells 
      attaching to derivatized SAMs. Well-controlled alkylthiol surface chemistries 
      were used to monitor and modulate the activation state of RhoA in attaching 
      cells. Activation states were determined indirectly by fractionating cell lysates 
      into membrane and cytosolic fractions by ultracentrifugation. Western blots were 
      then performed, showing RhoA localization to be surface chemistry-dependent. 
      RhoGDI levels and its intracellular localization were also shown to be 
      surface-chemistry dependent. Cells cultured on -CH3 terminated SAMs, which 
      normally exhibit a low-growth phenotype, were transfected with a constitutively 
      active mutant form of RhoA. Subsequent cell morphological changes were observed 
      on SAM surfaces by fluorescence microscopy. Results support surface chemistry 
      influences on the activation state of RhoA mediated by adsorbed proteins and 
      distinct changes in adherent cell morphology resulting from modulation of this 
      activation state.
FAU - McClary, K B
AU  - McClary KB
AD  - Department of Biochemistry, Colorado State University, Ft. Collins 80523-1872, 
      USA.
FAU - Grainger, D W
AU  - Grainger DW
LA  - eng
GR  - R01 GM56751-01/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - Netherlands
TA  - Biomaterials
JT  - Biomaterials
JID - 8100316
RN  - 0 (Fibronectins)
RN  - EC 2.4.2.- (ADP Ribose Transferases)
RN  - EC 2.4.2.- (exoenzyme C3, Clostridium botulinum)
RN  - EC 3.4.24.69 (Botulinum Toxins)
RN  - EC 3.6.5.2 (rhoA GTP-Binding Protein)
SB  - IM
MH  - 3T3 Cells
MH  - ADP Ribose Transferases/metabolism
MH  - Animals
MH  - Blotting, Western
MH  - *Botulinum Toxins
MH  - Cell Division
MH  - Cell Size
MH  - Cytosol/metabolism
MH  - Fibroblasts/*metabolism
MH  - Fibronectins/metabolism
MH  - Mice
MH  - Signal Transduction
MH  - Surface Properties
MH  - Time Factors
MH  - rhoA GTP-Binding Protein/*metabolism
EDAT- 1999/12/30 09:00
MHDA- 2000/02/19 09:00
CRDT- 1999/12/30 09:00
PHST- 1999/12/30 09:00 [pubmed]
PHST- 2000/02/19 09:00 [medline]
PHST- 1999/12/30 09:00 [entrez]
AID - S0142-9612(99)00171-4 [pii]
AID - 10.1016/s0142-9612(99)00171-4 [doi]
PST - ppublish
SO  - Biomaterials. 1999 Dec;20(23-24):2435-46. doi: 10.1016/s0142-9612(99)00171-4.