Different Susceptibility of Protein Kinases to Staurosporine Inhibition

Meggio, Flavio; Deana, Arianna Donella; Ruzzene, Maria; Brunati, Anna Maria; Cesaro, Luca; Guerra, Bárbara; Meyer, Thomas; Mett, Helmut; Fabbro, Doriano; Furet, Pascal; Dobrowolska, Grażyna; Pinna, Lorenzo A.

doi:10.1111/j.1432-1033.1995.317_c.x

Cited by 268 publications

(197 citation statements)

References 32 publications

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“…Neither cAMP-dependent protein kinase nor protein kinase C were capable of phosphorylating the hinge regions. Furthermore, the AP-3 kinase activity was insensitive to staurosporine, a promiscuous inhibitor of serine-threonine kinases (Meggio et al, 1995). The casein kinases were attractive candidates because they prefer to phosphorylate near acidic amino acids (Tuazon and Traugh, 1991), which are rich in the ␤3 hinge region, because of the genetic interactions in yeast between AP-3 and casein kinase (Panek et al, 1997) and because casein kinases are insensitive to staurosporine (Meggio et al,1995).…”

Section: The Ap-3-associated Kinase Activitymentioning

confidence: 99%

Section: ␤3 Subunit Of the Ap-3 Complex Is Thiophosphorylatedmentioning

confidence: 99%

“…We compared the effects of a broad-spectrum inhibitor of serine-threonine kinases, staurosporine (Meggio et al, 1995), to the selective casein kinase inhibitor CKI-7 (Chijiwa et al, 1989). CKI-7 was studied since genetic interactions between AP-3 complex subunits and casein kinase I genes have been The hinge domain of ␤3A and ␤3B are selectively thiophosphorylated by casein kinase I. GST and GST fusion proteins encompassing the trunk of ␤3A (residues 1-287 and 288 -642; lanes 2 and 3, respectively), the hinge domain of ␤3A (residues 643-809; lanes 4, 7, 11, and 15) and ␤3B (residues 647-796; lanes 8, 12, and 16), the ear domain of ␤3A (residues 810 -1094, lane 5), or the hinge-COOH terminal region of ␤2 (residues 592-951; lanes 9, 13, and 17) were thiophosphorylated as described with purified casein kinase I (10 U/assay; lanes 1-9), catalytic subunit of the cAMP-dependent kinase (10U/assay; lanes 10 -13), or the catalytic tryptic fragment of brain protein kinase C (20 ng/assay; lanes 14 -17).…”

Section: ␤3 Subunit Of the Ap-3 Complex Is Thiophosphorylatedmentioning

confidence: 99%

“…11, August 2000 2595 reported in yeast (Panek et al, 1997). The presence of the thiophosphorylated ␤3 subunit was decreased by CKI-7 or heparin, another inhibitor of casein kinases (Zhai et al, 1995) ( Figure 2b, lanes 2 and 3, 6 and 7) but not by high concentrations of staurosporine (Figure 2b, lanes 4 and 8), which is unable to inhibit casein kinases (Meggio et al, 1995). Identical results were obtained whether the total proteins were recruited to synaptic vesicles ( Figure 2b, lanes 1-4) or the immunoprecipitated ␤3 subunit were analyzed (Figure 2b, lanes 5-8).…”

mentioning

confidence: 99%

“…The 150-kDa band could be immunoprecipitated by antibodies against the ␤3 subunit of the AP-3 complex (Figure 2b, lane 5). No other thiophosphorylated bands were apparent after solubilizing the coated synaptic vesicles.We compared the effects of a broad-spectrum inhibitor of serine-threonine kinases, staurosporine (Meggio et al, 1995), to the selective casein kinase inhibitor CKI-7 (Chijiwa et al, 1989). CKI-7 was studied since genetic interactions between AP-3 complex subunits and casein kinase I genes have been Figure 3.…”

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The AP-3 Complex Required for Endosomal Synaptic Vesicle Biogenesis is Associated with a Casein Kinase Ια-Like Isoform

Faúndez

Kelly

2000

MBoC

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The formation of small vesicles is mediated by cytoplasmic coats the assembly of which is regulated by the activity of GTPases, kinases, and phosphatases. A heterotetrameric AP-3 adaptor complex has been implicated in the formation of synaptic vesicles from PC12 endosomes . When the small GTPase ARF1 is prevented from hydrolyzing GTP, we can reconstitute AP-3 recruitment to synaptic vesicle membranes in an assembly reaction that requires temperatures above 15°C and the presence of ATP suggesting that an enzymatic step is involved in the coat assembly. We have now found an enzymatic reaction, the phosphorylation of the AP-3 adaptor complex, that is linked with synaptic vesicle coating. Phosphorylation occurs in the ␤3 subunit of the complex by a kinase similar to casein kinase 1␣. The kinase copurifies with neuronal-specific AP-3. In vitro, purified casein kinase I selectively phosphorylates the ␤3A and ␤3B subunit at its hinge domain. Inhibiting the kinase hinders the recruitment of AP-3 to synaptic vesicles. The same inhibitors that prevent coat assembly in vitro also inhibit the formation of synaptic vesicles in PC12 cells. The data suggest, therefore, that the mechanism of AP-3-mediated vesiculation from neuroendocrine endosomes requires the phosphorylation of the adaptor complex at a step during or after AP-3 recruitment to membranes. INTRODUCTIONMembrane proteins are carried from donor membranes to acceptor membranes by two steps, the vesiculation of a carrier vesicle from the donor and the fusion of the carrier vesicle with the acceptor. Vesiculation of the donor membrane can itself be divided into five steps: recognition of cargo proteins; recruitment of coating molecules; the imposition of curvature on the forming vesicle membrane; fission of the carrier vesicle from the donor membrane; and, finally, the loss of the vesicle coat (Springer et al., 1999). To discover the molecular events that take place at each step, several laboratories have reconstituted either the entire process of vesiculation or individual steps in vitro.The first two steps of vesiculation, cargo recognition and coating, can be regulated by phosphorylation. Membrane proteins are recognized as cargo because they contain sorting domains. Adaptors interact with a tyrosine or a dileucine motif (Schmid, 1997;Kirchhausen et al., 1997;Bonifacino and Dell'Angelica, 1999). The recognition of cargo molecules can be regulated by the phosphorylation of sites close to the sorting domains; for example, in the trafficking of furin (Wan et al., 1998;Teuchert et al., 1999;Molloy et al., 1999), CD4 (Pitcher et al., 1999), CTLA-4 (Shiratori et al., 1997), MHC-II-Ii complex (Anderson and Roche, 1998), and pIgAR (Casanova et al., 1990;Okamoto et al., 1994;Luton et al., 1998). The cargo molecules are recruited into a newly forming carrier vesicle by cytoplasmic coat molecules (Matsuoka et al., 1998;Bremser et al., 1999). The recruitment of two of the known coats, COPI and COPII, is regulated by small ARF-like GTPases and appears to be a relatively simple p...

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Section: The Ap-3-associated Kinase Activitymentioning

confidence: 99%

Section: ␤3 Subunit Of the Ap-3 Complex Is Thiophosphorylatedmentioning

confidence: 99%

Section: ␤3 Subunit Of the Ap-3 Complex Is Thiophosphorylatedmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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The AP-3 Complex Required for Endosomal Synaptic Vesicle Biogenesis is Associated with a Casein Kinase Ια-Like Isoform

Faúndez

Kelly

2000

MBoC

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How to bring orphan genes into functional families

Bianchi

Sartori

Vandenbol

et al. 1999

Yeast

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In the framework of the B1 Consortium of the EUROFAN‐1 project, we set up a series of simple phenotypic tests that can be performed on a large number of strains at a time. This methodological approach was intended to help assign functions of putative genes coding for unknown proteins to several specific aspects of cell biology. The tests were chosen to study phenotypes which should be affected by numerous genes. In this report, we examined the sensitivity/resistance or the adaptation of the cell to physical or chemical stresses (thermotolerance, osmotolerance and ethanol sensitivity), the effects of the alteration of the level of protein phosphorylation (sensitivity or resistance to compounds affecting the activity of protein kinases or phosphatases) and the effects of compounds interfering with synthesis of nucleic acids or proteins. Deletions in 66 genes of unknown function have been tested in 21 different conditions. In many deletant strains, phenotypes were observed and, for the most promising candidates, tetrad analysis was performed in order to verify co‐segregation of the deletion marker with the phenotype. Copyright © 1999 John Wiley & Sons, Ltd.

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How to bring orphan genes into functional families

Bianchi¹,

Sartori²,

Vandenbol³

et al. 1999

Yeast

View full text Add to dashboard Cite

In the framework of the B1 Consortium of the EUROFAN-1 project, we set up a series of simple phenotypic tests that can be performed on a large number of strains at a time. This methodological approach was intended to help assign functions of putative genes coding for unknown proteins to several specific aspects of cell biology. The tests were chosen to study phenotypes which should be affected by numerous genes. In this report, we examined the sensitivity/resistance or the adaptation of the cell to physical or chemical stresses (thermotolerance, osmotolerance and ethanol sensitivity), the effects of the alteration of the level of protein phosphorylation (sensitivity or resistance to compounds affecting the activity of protein kinases or phosphatases) and the effects of compounds interfering with synthesis of nucleic acids or proteins. Deletions in 66 genes of unknown function have been tested in 21 different conditions. In many deletant strains, phenotypes were observed and, for the most promising candidates, tetrad analysis was performed in order to verify co-segregation of the deletion marker with the phenotype.

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Different Susceptibility of Protein Kinases to Staurosporine Inhibition

Cited by 268 publications

References 32 publications

The AP-3 Complex Required for Endosomal Synaptic Vesicle Biogenesis is Associated with a Casein Kinase Ια-Like Isoform

The AP-3 Complex Required for Endosomal Synaptic Vesicle Biogenesis is Associated with a Casein Kinase Ια-Like Isoform

How to bring orphan genes into functional families

How to bring orphan genes into functional families

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