Frameshift mutation S368fs in the gene encoding cytoskeletal β-actin leads to ACTB-associated syndromic thrombocytopenia by impairing actin dynamics
Heterozygous dominant mutations in the ubiquitously produced cytoskeletal β–actin isoform lead to a broad range of human disease phenotypes, which are currently classified as three distinct clinical entities termed Baraitser-Winter–Cerebrofrontofacial syndrome (BWCFF), ACTB–associated pleiotropic ma...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-04-01
|
| Series: | European Journal of Cell Biology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S017193352200019X |
| _version_ | 1818217542081052672 |
|---|---|
| author | Johannes N. Greve Frederic V. Schwäbe Thomas Pokrant Jan Faix Nataliya Di Donato Manuel H. Taft Dietmar J. Manstein |
| author_facet | Johannes N. Greve Frederic V. Schwäbe Thomas Pokrant Jan Faix Nataliya Di Donato Manuel H. Taft Dietmar J. Manstein |
| author_sort | Johannes N. Greve |
| collection | DOAJ |
| description | Heterozygous dominant mutations in the ubiquitously produced cytoskeletal β–actin isoform lead to a broad range of human disease phenotypes, which are currently classified as three distinct clinical entities termed Baraitser-Winter–Cerebrofrontofacial syndrome (BWCFF), ACTB–associated pleiotropic malformation syndrome with intellectual disability (ACTB–PMSID), and ACTB–associated syndromic thrombocytopenia (ACTB–AST). The latter two are distinguishable from BWCFF by the presence of milder craniofacial features and less pronounced developmental abnormalities, or the absence of craniofacial features in combination with a characteristic thrombocytopenia with platelet anisotropy. Production and correct function of β–actin is required for multiple essential processes in all types of cells. Directed cell migration, cytokinesis and morphogenesis are amongst the functions that are supported by β–actin. Here we report the recombinant production and biochemical characterization of the ACTB–AST mutant p.S368fs, resulting in an altered sequence in the C–terminal region of β–actin that includes a replacement of the last 8 residues and an elongation of the molecule by 4 residues. The mutation affects a region important for actin polymerization and actin–profilin interaction. Accordingly, we measured markedly reduced rates of nucleation and polymerization during spontaneous actin assembly and lower affinity of p.S368fs for human profilin–1. The reduced affinity is also reflected in the lower propensity of profilin–1 to extend the nucleation phase of p.S368fs. While localized in close proximity to actin–cofilin and actin–myosin interfaces, we determined only minor effects of the mutation on the interaction of mutant filaments with cofilin and myosin family members. However, allosteric effects on sites distant from the mutation manifest themselves in a 7.9 °C reduction in thermal denaturation temperature, a 2–fold increase in the observed IC50 for DNase–I, and changes in nucleotide exchange kinetics. Our results support a disease mechanism involving impaired actin dynamics and function through disruption of actin–profilin interactions and further exacerbated by allosteric perturbations. |
| first_indexed | 2024-12-12T07:09:31Z |
| format | Article |
| id | doaj.art-f4cd10b5d3b14de5b63c994899b160c7 |
| institution | Directory Open Access Journal |
| issn | 0171-9335 |
| language | English |
| last_indexed | 2024-12-12T07:09:31Z |
| publishDate | 2022-04-01 |
| publisher | Elsevier |
| record_format | Article |
| series | European Journal of Cell Biology |
| spelling | doaj.art-f4cd10b5d3b14de5b63c994899b160c72022-12-22T00:33:39ZengElsevierEuropean Journal of Cell Biology0171-93352022-04-011012151216Frameshift mutation S368fs in the gene encoding cytoskeletal β-actin leads to ACTB-associated syndromic thrombocytopenia by impairing actin dynamicsJohannes N. Greve0Frederic V. Schwäbe1Thomas Pokrant2Jan Faix3Nataliya Di Donato4Manuel H. Taft5Dietmar J. Manstein6Institute for Biophysical Chemistry and Structural Biochemistry, Medizinische Hochschule Hannover, Hannover 30625, GermanyInstitute for Biophysical Chemistry and Structural Biochemistry, Medizinische Hochschule Hannover, Hannover 30625, GermanyInstitute for Biophysical Chemistry and Structural Biochemistry, Medizinische Hochschule Hannover, Hannover 30625, GermanyInstitute for Biophysical Chemistry and Structural Biochemistry, Medizinische Hochschule Hannover, Hannover 30625, GermanyInstitute for Clinical Genetics, University Hospital, TU Dresden, Dresden 01307, GermanyInstitute for Biophysical Chemistry and Structural Biochemistry, Medizinische Hochschule Hannover, Hannover 30625, GermanyInstitute for Biophysical Chemistry and Structural Biochemistry, Medizinische Hochschule Hannover, Hannover 30625, Germany; Division for Structural Biochemistry, Hannover Medical School, Carl Neuberg Str. 1, D–30625 Hannover, Germany; RESiST, Cluster of Excellence 2155, Medizinische Hochschule Hannover, 30625 Hannover, Germany; Correspondence to: Institute for Biophysical Chemistry, Medizinische Hochschule Hannover, Fritz Hartmann Centre for Medical Research, Carl-Neuberg-Str. 1, 30625 Hannover, Germany.Heterozygous dominant mutations in the ubiquitously produced cytoskeletal β–actin isoform lead to a broad range of human disease phenotypes, which are currently classified as three distinct clinical entities termed Baraitser-Winter–Cerebrofrontofacial syndrome (BWCFF), ACTB–associated pleiotropic malformation syndrome with intellectual disability (ACTB–PMSID), and ACTB–associated syndromic thrombocytopenia (ACTB–AST). The latter two are distinguishable from BWCFF by the presence of milder craniofacial features and less pronounced developmental abnormalities, or the absence of craniofacial features in combination with a characteristic thrombocytopenia with platelet anisotropy. Production and correct function of β–actin is required for multiple essential processes in all types of cells. Directed cell migration, cytokinesis and morphogenesis are amongst the functions that are supported by β–actin. Here we report the recombinant production and biochemical characterization of the ACTB–AST mutant p.S368fs, resulting in an altered sequence in the C–terminal region of β–actin that includes a replacement of the last 8 residues and an elongation of the molecule by 4 residues. The mutation affects a region important for actin polymerization and actin–profilin interaction. Accordingly, we measured markedly reduced rates of nucleation and polymerization during spontaneous actin assembly and lower affinity of p.S368fs for human profilin–1. The reduced affinity is also reflected in the lower propensity of profilin–1 to extend the nucleation phase of p.S368fs. While localized in close proximity to actin–cofilin and actin–myosin interfaces, we determined only minor effects of the mutation on the interaction of mutant filaments with cofilin and myosin family members. However, allosteric effects on sites distant from the mutation manifest themselves in a 7.9 °C reduction in thermal denaturation temperature, a 2–fold increase in the observed IC50 for DNase–I, and changes in nucleotide exchange kinetics. Our results support a disease mechanism involving impaired actin dynamics and function through disruption of actin–profilin interactions and further exacerbated by allosteric perturbations.http://www.sciencedirect.com/science/article/pii/S017193352200019XActinopathyAllosteryCofilinCytoskeletonMyosinProfilin |
| spellingShingle | Johannes N. Greve Frederic V. Schwäbe Thomas Pokrant Jan Faix Nataliya Di Donato Manuel H. Taft Dietmar J. Manstein Frameshift mutation S368fs in the gene encoding cytoskeletal β-actin leads to ACTB-associated syndromic thrombocytopenia by impairing actin dynamics European Journal of Cell Biology Actinopathy Allostery Cofilin Cytoskeleton Myosin Profilin |
| title | Frameshift mutation S368fs in the gene encoding cytoskeletal β-actin leads to ACTB-associated syndromic thrombocytopenia by impairing actin dynamics |
| title_full | Frameshift mutation S368fs in the gene encoding cytoskeletal β-actin leads to ACTB-associated syndromic thrombocytopenia by impairing actin dynamics |
| title_fullStr | Frameshift mutation S368fs in the gene encoding cytoskeletal β-actin leads to ACTB-associated syndromic thrombocytopenia by impairing actin dynamics |
| title_full_unstemmed | Frameshift mutation S368fs in the gene encoding cytoskeletal β-actin leads to ACTB-associated syndromic thrombocytopenia by impairing actin dynamics |
| title_short | Frameshift mutation S368fs in the gene encoding cytoskeletal β-actin leads to ACTB-associated syndromic thrombocytopenia by impairing actin dynamics |
| title_sort | frameshift mutation s368fs in the gene encoding cytoskeletal β actin leads to actb associated syndromic thrombocytopenia by impairing actin dynamics |
| topic | Actinopathy Allostery Cofilin Cytoskeleton Myosin Profilin |
| url | http://www.sciencedirect.com/science/article/pii/S017193352200019X |
| work_keys_str_mv | AT johannesngreve frameshiftmutations368fsinthegeneencodingcytoskeletalbactinleadstoactbassociatedsyndromicthrombocytopeniabyimpairingactindynamics AT fredericvschwabe frameshiftmutations368fsinthegeneencodingcytoskeletalbactinleadstoactbassociatedsyndromicthrombocytopeniabyimpairingactindynamics AT thomaspokrant frameshiftmutations368fsinthegeneencodingcytoskeletalbactinleadstoactbassociatedsyndromicthrombocytopeniabyimpairingactindynamics AT janfaix frameshiftmutations368fsinthegeneencodingcytoskeletalbactinleadstoactbassociatedsyndromicthrombocytopeniabyimpairingactindynamics AT nataliyadidonato frameshiftmutations368fsinthegeneencodingcytoskeletalbactinleadstoactbassociatedsyndromicthrombocytopeniabyimpairingactindynamics AT manuelhtaft frameshiftmutations368fsinthegeneencodingcytoskeletalbactinleadstoactbassociatedsyndromicthrombocytopeniabyimpairingactindynamics AT dietmarjmanstein frameshiftmutations368fsinthegeneencodingcytoskeletalbactinleadstoactbassociatedsyndromicthrombocytopeniabyimpairingactindynamics |