Abstract
ALTHOUGH vinculin is present at all sites of F-actin attachment to plasma membranes1 and is required for linkage of myofibrils to sarcolemma2, it is unclear how it promotes attachment of actin to membranes. Because biochemical evidence for a direct interaction of vinculin with F-actin is controversial3-9, current models of actin-membrane linkages depict only an indirect role for vinculin, as a tether for α-actinin10. We demonstrate here that an intramolecular association between the 95K head and 30K tail domains of vinculin11 masks an F-actin binding site present in the carboxy-terminal tail domain. Cosedimentation and crosslinking assays, and direct visualization by transmission electron microscopy, reveal an interaction between F-actin and a bacterially expressed fusion protein containing amino acids 811-1066 of vinculin, and between F-actin and a proteolytic fragment of vinculin containing amino acids 858-1066. Vinculin itself neither cosediments with nor crosslinks F-actin. The amino-terminal 95K head fragment of vinculin, but not intact vinculin, inhibits both cosedimentation and crosslinking. We propose that assembly of vinculin into an adherens junction involves disruption of the head-tail interaction, revealing a site that mediates microfilament attachment.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 261-264 |
| Number of pages | 4 |
| Journal | Nature |
| Volume | 373 |
| Issue number | 6511 |
| State | Published - 1995 |
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Cite this
F-actin binding site masked by the intramolecular association of vinculin head and tail domains. / Johnson, Robert P.; Craig, Susan W.
In: Nature, Vol. 373, No. 6511, 1995, p. 261-264.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - F-actin binding site masked by the intramolecular association of vinculin head and tail domains
AU - Johnson, Robert P.
AU - Craig, Susan W.
PY - 1995
Y1 - 1995
N2 - ALTHOUGH vinculin is present at all sites of F-actin attachment to plasma membranes1 and is required for linkage of myofibrils to sarcolemma2, it is unclear how it promotes attachment of actin to membranes. Because biochemical evidence for a direct interaction of vinculin with F-actin is controversial3-9, current models of actin-membrane linkages depict only an indirect role for vinculin, as a tether for α-actinin10. We demonstrate here that an intramolecular association between the 95K head and 30K tail domains of vinculin11 masks an F-actin binding site present in the carboxy-terminal tail domain. Cosedimentation and crosslinking assays, and direct visualization by transmission electron microscopy, reveal an interaction between F-actin and a bacterially expressed fusion protein containing amino acids 811-1066 of vinculin, and between F-actin and a proteolytic fragment of vinculin containing amino acids 858-1066. Vinculin itself neither cosediments with nor crosslinks F-actin. The amino-terminal 95K head fragment of vinculin, but not intact vinculin, inhibits both cosedimentation and crosslinking. We propose that assembly of vinculin into an adherens junction involves disruption of the head-tail interaction, revealing a site that mediates microfilament attachment.
AB - ALTHOUGH vinculin is present at all sites of F-actin attachment to plasma membranes1 and is required for linkage of myofibrils to sarcolemma2, it is unclear how it promotes attachment of actin to membranes. Because biochemical evidence for a direct interaction of vinculin with F-actin is controversial3-9, current models of actin-membrane linkages depict only an indirect role for vinculin, as a tether for α-actinin10. We demonstrate here that an intramolecular association between the 95K head and 30K tail domains of vinculin11 masks an F-actin binding site present in the carboxy-terminal tail domain. Cosedimentation and crosslinking assays, and direct visualization by transmission electron microscopy, reveal an interaction between F-actin and a bacterially expressed fusion protein containing amino acids 811-1066 of vinculin, and between F-actin and a proteolytic fragment of vinculin containing amino acids 858-1066. Vinculin itself neither cosediments with nor crosslinks F-actin. The amino-terminal 95K head fragment of vinculin, but not intact vinculin, inhibits both cosedimentation and crosslinking. We propose that assembly of vinculin into an adherens junction involves disruption of the head-tail interaction, revealing a site that mediates microfilament attachment.
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UR - http://www.scopus.com/inward/citedby.url?scp=0028836195&partnerID=8YFLogxK
M3 - Article
C2 - 7816144
AN - SCOPUS:0028836195
VL - 373
SP - 261
EP - 264
JO - Nature
JF - Nature
SN - 0028-0836
IS - 6511
ER -