The PKA regulatory subunit from yeast forms a homotetramer: Low-resolution structure of the N-terminal oligomerization domain
The cAMP dependent protein kinase (PKA) is a key enzyme involved in many cellular processes in eukaryotes. In mammals, the regulatory (R) subunit localises the catalytic (C) subunit to specific subcellular sites through the interaction of its N-terminal homodimeric docking and dimerization (D/D) dom...
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| Publicado: |
2016
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10478477_v193_n2_p141_GonzalezBardeci http://hdl.handle.net/20.500.12110/paper_10478477_v193_n2_p141_GonzalezBardeci |
| Aporte de: |
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paper:paper_10478477_v193_n2_p141_GonzalezBardeci |
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| record_format |
dspace |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Bcy1 D/D domain Protein kinase A Regulatory subunit Saccharomyces cerevisiae Bcy1 protein cyclic AMP dependent protein kinase homodimer tetramer unclassified drug Bcy1 protein, S cerevisiae cyclic AMP dependent protein kinase protein subunit Saccharomyces cerevisiae protein ab initio calculation amino terminal sequence Article circular dichroism cross linking docking and dimerization domain hydrodynamics light scattering molecular docking nonhuman oligomerization priority journal protein domain protein secondary structure Saccharomyces cerevisiae sedimentation size exclusion chromatography X ray crystallography chemistry genetics metabolism molecular model protein multimerization protein subunit protein tertiary structure small angle scattering X ray diffraction Chromatography, Gel Circular Dichroism Cyclic AMP-Dependent Protein Kinases Models, Molecular Protein Multimerization Protein Structure, Tertiary Protein Subunits Saccharomyces cerevisiae Proteins Scattering, Small Angle X-Ray Diffraction |
| spellingShingle |
Bcy1 D/D domain Protein kinase A Regulatory subunit Saccharomyces cerevisiae Bcy1 protein cyclic AMP dependent protein kinase homodimer tetramer unclassified drug Bcy1 protein, S cerevisiae cyclic AMP dependent protein kinase protein subunit Saccharomyces cerevisiae protein ab initio calculation amino terminal sequence Article circular dichroism cross linking docking and dimerization domain hydrodynamics light scattering molecular docking nonhuman oligomerization priority journal protein domain protein secondary structure Saccharomyces cerevisiae sedimentation size exclusion chromatography X ray crystallography chemistry genetics metabolism molecular model protein multimerization protein subunit protein tertiary structure small angle scattering X ray diffraction Chromatography, Gel Circular Dichroism Cyclic AMP-Dependent Protein Kinases Models, Molecular Protein Multimerization Protein Structure, Tertiary Protein Subunits Saccharomyces cerevisiae Proteins Scattering, Small Angle X-Ray Diffraction Rinaldi, Jimena Julieta Rossi, Silvia Graciela Moreno, Silvia The PKA regulatory subunit from yeast forms a homotetramer: Low-resolution structure of the N-terminal oligomerization domain |
| topic_facet |
Bcy1 D/D domain Protein kinase A Regulatory subunit Saccharomyces cerevisiae Bcy1 protein cyclic AMP dependent protein kinase homodimer tetramer unclassified drug Bcy1 protein, S cerevisiae cyclic AMP dependent protein kinase protein subunit Saccharomyces cerevisiae protein ab initio calculation amino terminal sequence Article circular dichroism cross linking docking and dimerization domain hydrodynamics light scattering molecular docking nonhuman oligomerization priority journal protein domain protein secondary structure Saccharomyces cerevisiae sedimentation size exclusion chromatography X ray crystallography chemistry genetics metabolism molecular model protein multimerization protein subunit protein tertiary structure small angle scattering X ray diffraction Chromatography, Gel Circular Dichroism Cyclic AMP-Dependent Protein Kinases Models, Molecular Protein Multimerization Protein Structure, Tertiary Protein Subunits Saccharomyces cerevisiae Proteins Scattering, Small Angle X-Ray Diffraction |
| description |
The cAMP dependent protein kinase (PKA) is a key enzyme involved in many cellular processes in eukaryotes. In mammals, the regulatory (R) subunit localises the catalytic (C) subunit to specific subcellular sites through the interaction of its N-terminal homodimeric docking and dimerization (D/D) domain with specific scaffold proteins. The structure of the D/D domain has been extensively studied in mammals, but there is little information from non-mammalian species. In this work, we present the structural analysis of the D/D domain of Bcy1, the R subunit of PKA from Saccharomyces cerevisiae. Using chemical crosslinking experiments and static light scattering measurements we found that this R subunit forms a tetramer in solution, unlike its dimeric mammalian counterparts. We determined that the D/D domain is responsible for this unusual oligomeric state. Using biophysical techniques including size-exclusion chromatography, sucrose gradient sedimentation, small angle X-ray scattering (SAXS), and circular dichroism, we performed a detailed structural characterization of the tetrameric D/D domain of Bcy1. We used homology modelling in combination with computer-aided docking methods and ab initio SAXS modelling methods to develop structural models for the D/D domain tetramer. The models consist of two homodimers with a canonical D/D domain fold that generate a dimer of dimers with novel putative interaction surfaces. These findings indicate that the oligomerization states of PKA R subunits is more diverse than previously thought, and suggest that this might allow some forms of PKA to interact with a wide range of intracellular partners. © 2015 Elsevier Inc. |
| author |
Rinaldi, Jimena Julieta Rossi, Silvia Graciela Moreno, Silvia |
| author_facet |
Rinaldi, Jimena Julieta Rossi, Silvia Graciela Moreno, Silvia |
| author_sort |
Rinaldi, Jimena Julieta |
| title |
The PKA regulatory subunit from yeast forms a homotetramer: Low-resolution structure of the N-terminal oligomerization domain |
| title_short |
The PKA regulatory subunit from yeast forms a homotetramer: Low-resolution structure of the N-terminal oligomerization domain |
| title_full |
The PKA regulatory subunit from yeast forms a homotetramer: Low-resolution structure of the N-terminal oligomerization domain |
| title_fullStr |
The PKA regulatory subunit from yeast forms a homotetramer: Low-resolution structure of the N-terminal oligomerization domain |
| title_full_unstemmed |
The PKA regulatory subunit from yeast forms a homotetramer: Low-resolution structure of the N-terminal oligomerization domain |
| title_sort |
pka regulatory subunit from yeast forms a homotetramer: low-resolution structure of the n-terminal oligomerization domain |
| publishDate |
2016 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_10478477_v193_n2_p141_GonzalezBardeci http://hdl.handle.net/20.500.12110/paper_10478477_v193_n2_p141_GonzalezBardeci |
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