PMID- 10508610
OWN - NLM
STAT- MEDLINE
DCOM- 20000214
LR  - 20191210
IS  - 0960-9822 (Print)
IS  - 0960-9822 (Linking)
VI  - 9
IP  - 18
DP  - 1999 Sep 23
TI  - Sema3A-induced growth-cone collapse is mediated by Rac1 amino acids 17-32.
PG  - 991-8
AB  - BACKGROUND: Neurons project their axons along specific pathways in order to
      establish appropriate connections with their target cells. The rate and direction
      of axonal growth is determined by interactions between the highly motile growth
      cone and environmental cues that can act in either an attractive or a repulsive
      manner. Locomotion is ultimately dependent upon the reorganisation of the actin
      cytoskeleton and an established role for the Rho family of small GTPases in
      regulating this process in non-neuronal cells identifies them as candidate
      signalling molecules in growth cones. An inactive form of Rac1 has recently been 
      shown to inhibit the 'growth-cone collapse' response induced by chick Sema3A, a
      protein that has recently been established as an important guidance cue. The
      molecular basis for this inhibition remains unclear. RESULTS: We have made a
      series of overlapping peptides from the amino-terminal region of Rac1 and
      rendered them cell permeable by synthesis in tandem with an established
      internalisation vector. We report here that a peptide encompassing Rac1 amino
      acids 17-32 binds directly to the established Rac1-interacting molecules PAK,
      WASP, 3BP-1 and p85beta(P13K), but not to p67(Phox). Furthermore, the peptide can
      compete with activated Rac1 for target binding, and inhibits Sema3A-induced
      growth-cone collapse. We also synthesised cell-permeable peptides that correspond
      to the Cdc42/Rac1-binding (CRIB) motifs present in PAK and N-WASP. Our results
      show that a CRIB-containing peptide from PAK, but not that from N-WASP, inhibits 
      growth-cone collapse and that the inhibitory activity correlates with binding to 
      Rac1 and not to Cdc42. CONCLUSIONS: Our results suggest that Sema3A-induced
      growth-cone collapse is mediated by Rac1 amino acids 17-32, and demonstrate the
      feasibility of designing new cell-permeable inhibitors of small GTPases.
FAU - Vastrik, I
AU  - Vastrik I
AD  - Molecular Neurobiology Group GKT School of Medicine King's College London London,
      SE1 9RT, Computational Genome Analysis Laboratory ICRF 44 Lincoln's Inn Fields,
      London, WC2A 3PX, UK.
FAU - Eickholt, B J
AU  - Eickholt BJ
FAU - Walsh, F S
AU  - Walsh FS
FAU - Ridley, A
AU  - Ridley A
FAU - Doherty, P
AU  - Doherty P
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Curr Biol
JT  - Current biology : CB
JID - 9107782
RN  - 0 (Carrier Proteins)
RN  - 0 (GTPase-Activating Proteins)
RN  - 0 (Glycoproteins)
RN  - 0 (Peptide Fragments)
RN  - 0 (Phosphoproteins)
RN  - 0 (Proteins)
RN  - 0 (Rac1 GTP-binding protein (17-32))
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Semaphorin-3A)
RN  - 0 (Sh3bp1 protein, mouse)
RN  - 0 (Wiskott-Aldrich Syndrome Protein)
RN  - 0 (neutrophil cytosol factor 67K)
RN  - EC 3.6.5.2 (cdc42 GTP-Binding Protein)
RN  - EC 3.6.5.2 (rac1 GTP-Binding Protein)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - COS Cells
MH  - Carrier Proteins/metabolism
MH  - Chick Embryo
MH  - Chlorocebus aethiops
MH  - *GTPase-Activating Proteins
MH  - Glycoproteins/*physiology
MH  - Growth Cones/drug effects/metabolism/*ultrastructure
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Peptide Fragments/chemical synthesis/*pharmacology
MH  - Phosphatidylinositol 3-Kinases/metabolism
MH  - Phosphoproteins/metabolism
MH  - Protein Conformation
MH  - Protein Structure, Tertiary
MH  - Proteins/metabolism
MH  - Recombinant Fusion Proteins/pharmacology
MH  - Semaphorin-3A
MH  - Sequence Alignment
MH  - Sequence Homology, Amino Acid
MH  - Wiskott-Aldrich Syndrome Protein
MH  - cdc42 GTP-Binding Protein/physiology
MH  - rac1 GTP-Binding Protein/chemical synthesis/chemistry/*pharmacology/*physiology
EDAT- 1999/10/06 09:00
MHDA- 2000/02/19 09:00
CRDT- 1999/10/06 09:00
PHST- 1999/10/06 09:00 [pubmed]
PHST- 2000/02/19 09:00 [medline]
PHST- 1999/10/06 09:00 [entrez]
AID - S0960-9822(99)80447-3 [pii]
AID - 10.1016/s0960-9822(99)80447-3 [doi]
PST - ppublish
SO  - Curr Biol. 1999 Sep 23;9(18):991-8. doi: 10.1016/s0960-9822(99)80447-3.