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N-terminal processing of beta and gamma actin in HAP1 cells

ABSTRACT: Actin is one of the most essential and abundant intracellular proteins, playing an essential physiological role as the major constituent of the actin cytoskeleton. Two cytoplasmic actins, beta- and gamma-actin, are encoded by different genes, but their amino acid sequences differ only by four conservative substitutions at the N-terminus, making it very difficult to dissect their individual regulation in vivo. The majority of actins are N-terminally acetylated, following the removal of N-terminal Met. Here, we analyzed beta and gamma cytoplasmic actin N-termini in different actin preparations.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Haploid Cell

SUBMITTER: Dawei Dong

LAB HEAD: Anna Kashina

PROVIDER: PXD031481 | Pride | 2023-03-11

REPOSITORIES: Pride

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			Action	DRS
	KashinaA-19-G152-D23ENPEP-KO-3-QEP.raw	Raw
	KashinaA-19-G152-D23ENPEP-KO-7-QEP.raw	Raw
	KashinaA-19-G152-D23ENPEP-KO-8-QEP.raw	Raw
	KashinaA-19-G152-DNPEP-KO-13-QEP.raw	Raw
	KashinaA-19-G152-DNPEP-KO-20-QEP.raw	Raw

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Publications

Differential N-terminal processing of beta and gamma actin.

Chen Li L Vedula Pavan P Tang Hsin Yao HY Dong Dawei W DW Kashina Anna S AS

iScience 20220923 10

Cytoplasmic beta- and gamma-actin are ubiquitously expressed in every eukaryotic cell. They are encoded by different genes, but their amino acid sequences differ only by four conservative substitutions at the N-termini, making it difficult to dissect their individual regulation. Here, we analyzed actin from cultured cells and tissues by mass spectrometry and found that beta, unlike gamma actin, undergoes sequential removal of N-terminal Asp residues, leading to truncated actin species found in b ...[more]

PMID: 36248738

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Actb knockout in primary mouse embryonic fibroblasts LC-MS/MS

Project description:The cytoplasmic actin proteins, beta- and gamma-actin, are 99% identical but perform non-redundant functions. Genes encoding the cytoplasmic actins, Actb and Actg1, respectively, are less similar but still share 89% of their nucleotide sequences. Knockout (KO) of Actb by deletion of first coding exons 2 and 3 in mice is embryonic lethal while KO embryonic fibroblasts (MEFs) fail to proliferate. In contrast, KO of Actg1 is viable but mice lacking Actg1 present with increased perinatal lethality and Actg1 KO MEFs present with a much milder defect in cell proliferation. Recent studies have identified important protein-independent functions for both Actb and Actg1 and demonstrate that deletions within the Actb nucleotide sequence, and not loss of the beta-actin protein, cause the most severe phenotypes in KO mice and cells. Here, we use a multi-omics approach to better understand what drives the phenotypes of Actb KO MEFs. RNA-sequencing and mass spectrometry of Actb KO MEFs reveal largescale changes to the transcriptome, proteome, and phosphoproteome. Pathway analysis of genes and proteins differentially expressed upon Actb KO shows widespread dysregulation of genes involved in the cell cycle. Together, these data suggest novel, protein-independent roles for Actb in regulating gene expression associated with control of cell proliferation.

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