Cytoarchitecture and cell growth control

Slack, Jill K.; Higgins, Paul J.

doi:10.1002/(sici)1097-0169(1996)33:2<83::aid-cm1>3.0.co;2-j

Cited by 9 publications

(2 citation statements)

References 22 publications

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“…Moreover, focal adhesion proteins, such as integrin, that span the plasma membrane, serve to connect the extracellular matrix and cytoplasmic actin cytoskeleton at focal adhesions during myogenesis. The cellular proliferation and gene reprogramming in response to cellular adhesion are likely to be mediated by integrins [Juliano and Haskill, 1993;Juliano and Varner, 1993;Miyamoto et al, 1995;Slack and Higgins, 1996]. Our results also suggested that FHL3 does not stay in the nucleus after C2C12 myogenesis but localized only at the focal adhesion plaques of myotubes.…”

Section: Discussionmentioning

confidence: 49%

Protein-protein interaction of FHL3 with FHL2 and visualization of their interaction by green fluorescent proteins (GFP) two-fusion fluorescence resonance energy transfer (FRET)

Lee

et al. 2000

J. Cell. Biochem.

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LIM domain proteins are found to be important regulators in cell growth, cell fate determination, cell differentiation and remodeling of the cell cytoskeleton. Human Four-and-a-half LIM-only protein 3 (FHL3) is a type of LIM-only protein that contains four tandemly repeated LIM motifs with an N-terminal single zinc finger (half LIM motif). FHL3 expresses predominantly in human skeletal muscle. In this report, FHL3 was shown to be a novel interacting partner of FHL2 using the yeast two-hybrid assay. Furthermore, site-directed mutagenesis of FHL3 indicated that the LIM2 of FHL3 is the essential LIM domain for interaction with FHL2. Green fluorescent protein (GFP) was used to tag FHL3 in order to study its distribution during myogenesis. Our result shows that FHL3 was localized in the focal adhesions and nucleus of the cells. FHL3 mainly stayed in the focal adhesion during myogenesis. Moreover, using site-directed mutagenesis, the LIM1 of FHL3 was identified as an essential LIM domain for its subcellular localization. Mutants of GFP have given rise to a novel technique, two-fusion fluorescence resonance energy transfer (FRET), in the determination of protein-protein interaction at particular subcellular locations of eukaryotic cells. To determine whether FHL2 and FHL3 can interact with one another and to locate the site of this interaction in a single intact mammalian cell, we fused FHL2 and FHL3 to different mutants of GFP and studied their interactions using FRET. BFP/GFP fusion constructs were cotransfected into muscle myoblast C2C12 to verify the colocalization and subcellular localization of FRET. We found that FHL2 and FHL3 were colocalized in the mitochondria of the C2C12 cells and FRET was observed by using an epi-fluorescent microscope equipped with an FRET specific filter set.

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Section: Discussionmentioning

confidence: 49%

Protein-protein interaction of FHL3 with FHL2 and visualization of their interaction by green fluorescent proteins (GFP) two-fusion fluorescence resonance energy transfer (FRET)

Lee

et al. 2000

J. Cell. Biochem.

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“…In addition, aggregated ECM receptors such as integrin that spans the plasma membrane, connect the extracellular matrix and cytoplasmic actin cytoskeleton at focal adhesions. The cellular proliferation and gene reprogramming in response to adhesion are likely to be mediated by integrins by uncertain mechanisms and transduction pathways [Juliano and Haskill, 1993;Juliano and Varner, 1993;Miyamoto et al, 1995;Slack and Higgins, 1996].…”

Section: Discussionmentioning

confidence: 99%

Translocation of a human focal adhesion LIM-only protein, FHL2, during myofibrillogenesis and identification of LIM2 as the principal determinants of FHL2 focal adhesion localization

Kotaka

Kostin

et al. 2000

Cell Motil. Cytoskeleton

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Attenuation of plasminogen activator inhibitor type-1 promoter activity in serum-stimulated renal epithelial cells by a distal 5? flanking region

Slack

Higgins

1999

Cell Motil. Cytoskeleton

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Urokinase plasminogen activator (u‐PA) and its fast acting type‐1 inhibitor (PAI‐1) localize to cellular focal adhesive structures and the ajoining proximal undersurface region, respectively (Kutz et al., J. Cell. Physiol. 176:8–18, 1997). PAI‐1 may function in this locale to modulate pericellular proteolytic activity, cell‐to‐substrate adhesion, or matrix‐dependent motility. While PAI‐1 synthesis is regulated in an immediate‐early response manner in growth “activated” renal cells coincident with cytoskeletal restructuring, adhesive influences both repress the amplitude and prolong the time course of serum‐induced PAI‐1 transcription (Ryan et al., Biochem. J. 314:1041–1046, 1996). To identify potential adhesion‐responsive elements within the PAI‐1 gene that function in this complex mode of expression control, reporter constructs containing defined directionally deleted PAI‐1 5′ genomic fragments cloned upstream of a CAT gene were employed in transient transfection assays. A 483‐bp distal PAI‐1 flanking segment (corresponding to nucleotides −2395 to −1912) conferred significant adhesion‐dependent attenuation on serum‐induced PAI‐1 transcription. This 483‐bp distal PAI‐1 segment functioned as a repressor of reporter (CAT) activity under both adhesive and suspension culture conditions, however, when ligated upstream of either an SV40 promoter/enhancer or a minimal PAI‐1 promoter. These data suggest that repressor elements located between −2395 and −1912 bp interact with more proximal adhesion‐dependent regulatory elements to affect PAI‐1 expression attenuation during serum stimulation of adherent renal epithelial cells. Cell Motil. Cytoskeleton 44:168–176, 1999. © 1999 Wiley‐Liss, Inc.

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Cytoarchitecture and cell growth control

Cited by 9 publications

References 22 publications

Protein-protein interaction of FHL3 with FHL2 and visualization of their interaction by green fluorescent proteins (GFP) two-fusion fluorescence resonance energy transfer (FRET)

Protein-protein interaction of FHL3 with FHL2 and visualization of their interaction by green fluorescent proteins (GFP) two-fusion fluorescence resonance energy transfer (FRET)

Translocation of a human focal adhesion LIM-only protein, FHL2, during myofibrillogenesis and identification of LIM2 as the principal determinants of FHL2 focal adhesion localization

Attenuation of plasminogen activator inhibitor type-1 promoter activity in serum-stimulated renal epithelial cells by a distal 5? flanking region

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