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Mutations in the Catalytic Loop HRD Motif Alter the Activity and Function of Drosophila Src64

The catalytic loop HRD motif is found in most protein kinases and these amino acids are predicted to perform functions in catalysis, transition to, and stabilization of the active conformation of the kinase domain. We have identified mutations in a Drosophila src gene, src64, that alter the three HR...

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Detalles Bibliográficos
Autores principales: Strong, Taylor C., Kaur, Gurvinder, Thomas, Jeffrey H.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3223231/
https://www.ncbi.nlm.nih.gov/pubmed/22132220
http://dx.doi.org/10.1371/journal.pone.0028100
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author Strong, Taylor C.
Kaur, Gurvinder
Thomas, Jeffrey H.
author_facet Strong, Taylor C.
Kaur, Gurvinder
Thomas, Jeffrey H.
author_sort Strong, Taylor C.
collection PubMed
description The catalytic loop HRD motif is found in most protein kinases and these amino acids are predicted to perform functions in catalysis, transition to, and stabilization of the active conformation of the kinase domain. We have identified mutations in a Drosophila src gene, src64, that alter the three HRD amino acids. We have analyzed the mutants for both biochemical activity and biological function during development. Mutation of the aspartate to asparagine eliminates biological function in cytoskeletal processes and severely reduces fertility, supporting the amino acid's critical role in enzymatic activity. The arginine to cysteine mutation has little to no effect on kinase activity or cytoskeletal reorganization, suggesting that the HRD arginine may not be critical for coordinating phosphotyrosine in the active conformation. The histidine to leucine mutant retains some kinase activity and biological function, suggesting that this amino acid may have a biochemical function in the active kinase that is independent of its side chain hydrogen bonding interactions in the active site. We also describe the phenotypic effects of other mutations in the SH2 and tyrosine kinase domains of src64, and we compare them to the phenotypic effects of the src64 null allele.
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spelling pubmed-32232312011-11-30 Mutations in the Catalytic Loop HRD Motif Alter the Activity and Function of Drosophila Src64 Strong, Taylor C. Kaur, Gurvinder Thomas, Jeffrey H. PLoS One Research Article The catalytic loop HRD motif is found in most protein kinases and these amino acids are predicted to perform functions in catalysis, transition to, and stabilization of the active conformation of the kinase domain. We have identified mutations in a Drosophila src gene, src64, that alter the three HRD amino acids. We have analyzed the mutants for both biochemical activity and biological function during development. Mutation of the aspartate to asparagine eliminates biological function in cytoskeletal processes and severely reduces fertility, supporting the amino acid's critical role in enzymatic activity. The arginine to cysteine mutation has little to no effect on kinase activity or cytoskeletal reorganization, suggesting that the HRD arginine may not be critical for coordinating phosphotyrosine in the active conformation. The histidine to leucine mutant retains some kinase activity and biological function, suggesting that this amino acid may have a biochemical function in the active kinase that is independent of its side chain hydrogen bonding interactions in the active site. We also describe the phenotypic effects of other mutations in the SH2 and tyrosine kinase domains of src64, and we compare them to the phenotypic effects of the src64 null allele. Public Library of Science 2011-11-23 /pmc/articles/PMC3223231/ /pubmed/22132220 http://dx.doi.org/10.1371/journal.pone.0028100 Text en Strong et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Strong, Taylor C.
Kaur, Gurvinder
Thomas, Jeffrey H.
Mutations in the Catalytic Loop HRD Motif Alter the Activity and Function of Drosophila Src64
title Mutations in the Catalytic Loop HRD Motif Alter the Activity and Function of Drosophila Src64
title_full Mutations in the Catalytic Loop HRD Motif Alter the Activity and Function of Drosophila Src64
title_fullStr Mutations in the Catalytic Loop HRD Motif Alter the Activity and Function of Drosophila Src64
title_full_unstemmed Mutations in the Catalytic Loop HRD Motif Alter the Activity and Function of Drosophila Src64
title_short Mutations in the Catalytic Loop HRD Motif Alter the Activity and Function of Drosophila Src64
title_sort mutations in the catalytic loop hrd motif alter the activity and function of drosophila src64
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3223231/
https://www.ncbi.nlm.nih.gov/pubmed/22132220
http://dx.doi.org/10.1371/journal.pone.0028100
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