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Kinesin delivers: identifying receptors for motor proteins.

Hollenbeck PJ - J. Cell Biol. (2001)

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907, USA. phollenb@purdue.edu

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GFP-tagged JIP-3 protein was expressed in CV-1 cells, where it colocalized with kinesin-I and with Golgi and early secretory vesicles, but not with mitochondria or the ER-to-Golgi intermediate compartment... When they probed the interaction of kinesin-I and JIP-3 by yeast two-hybrid analysis and coprecipitation methods, they found that the NH2-terminal domain of JIP-3 bound a region of the kinesin light chain (LC) that contains six tetratricopeptide repeat (TPR) motifs (Blatch and Lasle 1999)... They employed the kinesin LC TPR domain as bait in a yeast two-hybrid screen of a mouse brain cDNA library and fished out three binding partners for kinesin: not only JIP-3, but also JIP-1 and JIP-2, which are unrelated to JIP-3, but very similar to each other... But JIP-1 and JIP-2 resembled more closely other TPR-binding proteins, in that they interacted with kinesin via their COOH termini... Verhey et al. 2001 found that the mutation of a single tyrosine three residues from the COOH terminus eliminated JIP-1 binding to kinesin... So kinesin binds, but does it deliver? To address this, they examined the distribution JIP-1 in neuronal cell lines (Fig. 2) and found that the kinesin-I/JIP-1 interaction was necessary for JIP-1 to accumulate in the tips of the neurites... In addition, there is good evidence that some cargoes bind the kinesin LC, via other receptors, outside the TPR domain... For example, although several classes of organelles are thought to be moved by kinesin-I, Verhey et al. 2001 found that blocking the binding of the kinesin LC TPR domain to other proteins did not disrupt the organelle distribution in CAD cells... Also, antibody disruption of binding to the LC only displaces about one-third of the kinesin-I from vesicles (Yu et al. 1992), even when the antibodies are directed specifically against the TPR domain (Stenoien and Brady 1997)... Indeed, receptors or signaling molecules that bind to other kinesin family members have been reported already (Nagata et al. 1998; Nakagawa et al. 2000; Setou et al. 2000)... If you would like to take part in the construction of the phylogenetic tree of motor protein receptors, you had better order your PCR primers soon.

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Kinesin-I is a heterotetramer formed by a coiled-coil interaction between two heavy chains (HCs) and the binding of a LC to the COOH-terminal region of each HC. Each HC has an NH2-terminal catalytic motor domain that interacts with a microtubule during movement. The heterotetramer binds to its cargo at its tail, as shown. However, the exact nature of the interaction, what kinesin is binding to, and how, has not been clear. The LCs have been thought to be essential for this interaction, and the two recent papers discussed here (Bowman et al. 2000; Verhey et al. 2001) identify the TPR domain of the LC as a site of cargo binding. For illustrative purposes, the kinesin molecule is drawn here approximately three times larger than true scale relative to the microtubule and vesicle. (Figure courtesy of W.M. Saxton)
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Figure 1: Kinesin-I is a heterotetramer formed by a coiled-coil interaction between two heavy chains (HCs) and the binding of a LC to the COOH-terminal region of each HC. Each HC has an NH2-terminal catalytic motor domain that interacts with a microtubule during movement. The heterotetramer binds to its cargo at its tail, as shown. However, the exact nature of the interaction, what kinesin is binding to, and how, has not been clear. The LCs have been thought to be essential for this interaction, and the two recent papers discussed here (Bowman et al. 2000; Verhey et al. 2001) identify the TPR domain of the LC as a site of cargo binding. For illustrative purposes, the kinesin molecule is drawn here approximately three times larger than true scale relative to the microtubule and vesicle. (Figure courtesy of W.M. Saxton)

Mentions: In retrospect, it is not surprising that potential cargo receptors specifically bind the TPR domain of the kinesin LC. Kinesin-I is a heterotetramer comprised of 2 LCs and two heavy chains (HCs; see Fig. 1). It has been thought for some time that the kinesin tail region binds to the motor's cargo (Vale and Fletterick 1997). Both the HCs and LCs occupy this region of the tetramer, but the preponderance of genetic and biochemical data indicate that the LCs are important or even essential for cargo binding (Yu et al. 1992; Stenoien and Brady 1997; Gindhart et al. 1998; Tsai et al. 2000). Furthermore, the TPR domain of the LC stands out specifically as a likely binding site because antibodies directed against it disrupt kinesin–cargo interactions (Stenoien and Brady 1997), and it has well characterized, specific protein binding properties (Blatch and Lasle 1999).


Kinesin delivers: identifying receptors for motor proteins.

Hollenbeck PJ - J. Cell Biol. (2001)

Kinesin-I is a heterotetramer formed by a coiled-coil interaction between two heavy chains (HCs) and the binding of a LC to the COOH-terminal region of each HC. Each HC has an NH2-terminal catalytic motor domain that interacts with a microtubule during movement. The heterotetramer binds to its cargo at its tail, as shown. However, the exact nature of the interaction, what kinesin is binding to, and how, has not been clear. The LCs have been thought to be essential for this interaction, and the two recent papers discussed here (Bowman et al. 2000; Verhey et al. 2001) identify the TPR domain of the LC as a site of cargo binding. For illustrative purposes, the kinesin molecule is drawn here approximately three times larger than true scale relative to the microtubule and vesicle. (Figure courtesy of W.M. Saxton)
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2198802&req=5

Figure 1: Kinesin-I is a heterotetramer formed by a coiled-coil interaction between two heavy chains (HCs) and the binding of a LC to the COOH-terminal region of each HC. Each HC has an NH2-terminal catalytic motor domain that interacts with a microtubule during movement. The heterotetramer binds to its cargo at its tail, as shown. However, the exact nature of the interaction, what kinesin is binding to, and how, has not been clear. The LCs have been thought to be essential for this interaction, and the two recent papers discussed here (Bowman et al. 2000; Verhey et al. 2001) identify the TPR domain of the LC as a site of cargo binding. For illustrative purposes, the kinesin molecule is drawn here approximately three times larger than true scale relative to the microtubule and vesicle. (Figure courtesy of W.M. Saxton)
Mentions: In retrospect, it is not surprising that potential cargo receptors specifically bind the TPR domain of the kinesin LC. Kinesin-I is a heterotetramer comprised of 2 LCs and two heavy chains (HCs; see Fig. 1). It has been thought for some time that the kinesin tail region binds to the motor's cargo (Vale and Fletterick 1997). Both the HCs and LCs occupy this region of the tetramer, but the preponderance of genetic and biochemical data indicate that the LCs are important or even essential for cargo binding (Yu et al. 1992; Stenoien and Brady 1997; Gindhart et al. 1998; Tsai et al. 2000). Furthermore, the TPR domain of the LC stands out specifically as a likely binding site because antibodies directed against it disrupt kinesin–cargo interactions (Stenoien and Brady 1997), and it has well characterized, specific protein binding properties (Blatch and Lasle 1999).

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907, USA. phollenb@purdue.edu

AUTOMATICALLY GENERATED EXCERPT
Please rate it.

GFP-tagged JIP-3 protein was expressed in CV-1 cells, where it colocalized with kinesin-I and with Golgi and early secretory vesicles, but not with mitochondria or the ER-to-Golgi intermediate compartment... When they probed the interaction of kinesin-I and JIP-3 by yeast two-hybrid analysis and coprecipitation methods, they found that the NH2-terminal domain of JIP-3 bound a region of the kinesin light chain (LC) that contains six tetratricopeptide repeat (TPR) motifs (Blatch and Lasle 1999)... They employed the kinesin LC TPR domain as bait in a yeast two-hybrid screen of a mouse brain cDNA library and fished out three binding partners for kinesin: not only JIP-3, but also JIP-1 and JIP-2, which are unrelated to JIP-3, but very similar to each other... But JIP-1 and JIP-2 resembled more closely other TPR-binding proteins, in that they interacted with kinesin via their COOH termini... Verhey et al. 2001 found that the mutation of a single tyrosine three residues from the COOH terminus eliminated JIP-1 binding to kinesin... So kinesin binds, but does it deliver? To address this, they examined the distribution JIP-1 in neuronal cell lines (Fig. 2) and found that the kinesin-I/JIP-1 interaction was necessary for JIP-1 to accumulate in the tips of the neurites... In addition, there is good evidence that some cargoes bind the kinesin LC, via other receptors, outside the TPR domain... For example, although several classes of organelles are thought to be moved by kinesin-I, Verhey et al. 2001 found that blocking the binding of the kinesin LC TPR domain to other proteins did not disrupt the organelle distribution in CAD cells... Also, antibody disruption of binding to the LC only displaces about one-third of the kinesin-I from vesicles (Yu et al. 1992), even when the antibodies are directed specifically against the TPR domain (Stenoien and Brady 1997)... Indeed, receptors or signaling molecules that bind to other kinesin family members have been reported already (Nagata et al. 1998; Nakagawa et al. 2000; Setou et al. 2000)... If you would like to take part in the construction of the phylogenetic tree of motor protein receptors, you had better order your PCR primers soon.

Show MeSH
Related in: MedlinePlus