[Novel splicing isoform of actin-binding protein alpha-actinin 4 in epidermoid carcinoma cells A431]

Tsitologiia. 2012;54(1):25-32.
[Article in Russian]

Abstract

Alpha-actinin 4 (ACTN4) belongs to actin binding proteins of the spectrin superfamily. Structural organisation of actin fibres and focal contacts is considered to be its primary function in a cell. Besides that, nucleocytoplasmic shuffling of ACTN4 and its involvement in nuclear processes were demonstrated. Lately, additional isoforms of ACTN4 resulted from an alternative splicing has been described in various cell types and malignant tumours. In this study, we present investigation of a novel ACTN4 isoform of 80 kDa. The isoform was found in human epidermoid carcinoma cells A431, and it was not detected in human skin fibroblasts, normal human keratinocytes and transformed human embryonic cells HEK293T. Analysis of ACTN4 mRNA in A431 cells showed the presence of a splice variant that lacked the exons 2-8. The deleted exons code two calponin homology domains responsible for ACTN4 binding to F-actin. Intracellular distribution of the described ACTN4 isoform (ACTN4ISO) overexpressed in HEK293T cells differed from that of the full size protein. In the cytoplasm, ACTN4ISO was allocated diffusively with no colocalisation with actin cytoskeleton structures. Intranuclear distribution of ACTN4ISO also differed from that of the full size ACTN4. Nevertheless, immunochemical analysis demonstrated possibility of ACTN4ISO to form heterodimers with the full size protein. Additional investigations of novel isoform interactions with ACTN4 protein partners might clarify its functional features in A431 cells.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / metabolism
  • Actinin / genetics*
  • Actinin / metabolism
  • Actins / metabolism*
  • Alternative Splicing
  • Amino Acid Sequence / genetics*
  • Calcium-Binding Proteins / chemistry
  • Calcium-Binding Proteins / genetics
  • Calponins
  • Carcinoma, Squamous Cell / genetics*
  • Carcinoma, Squamous Cell / metabolism
  • Cell Line, Tumor
  • Cell Nucleus / metabolism
  • Cytoplasm / metabolism
  • Exons
  • Fibroblasts / cytology
  • Fibroblasts / metabolism
  • HEK293 Cells
  • Humans
  • Keratinocytes / cytology
  • Keratinocytes / metabolism
  • Microfilament Proteins / chemistry
  • Microfilament Proteins / genetics
  • Molecular Sequence Data
  • Protein Binding
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Protein Multimerization
  • RNA, Messenger / analysis
  • RNA, Messenger / biosynthesis*
  • Sequence Deletion / genetics*
  • Skin / cytology
  • Skin / metabolism

Substances

  • ACTN4 protein, human
  • Actins
  • Calcium-Binding Proteins
  • Microfilament Proteins
  • Protein Isoforms
  • RNA, Messenger
  • Actinin