Molecular cloning and characterization of rat LC3A and LC3B - Two novel markers of autophagosome

Jiaxue Wu, Yongjun Dang, Wei Su, Chao Liu, Haijie Ma, Yuxi Shan, Yuan Pei, Bo Wan, Jinhu Guo, Long Yu

Research output: Contribution to journalArticle

Abstract

Rat microtubule-associated protein light chain 3 (LC3) is a homologue of yeast Atg8, an essential component of autophagy. Following synthesis, the C-terminus of rat LC3 is cleaved by a cysteine protease-Atg4, to produce LC3-I, which is located in cytosolic fraction. LC3-I can be converted to LC3-II through the processing by Atg7 (E1-like enzyme) and Atg3 (E2-like enzyme). LC3-II is modified by phosphatidylethanolamine on C-terminus and binds tightly to autophagosomal membrane. Here we reported the cloning of two novel variants of rat LC3, named LC3A and LC3B, respectively, and LC3B is an alternative splicing variant of LC3. LC3A, LC3B, and LC3 showed different expression patterns in rat tissues, suggesting a functional divergence among these proteins. When LC3A and LC3B were overexpressed, both exhibited two forms (18 and 16 kDa, representing types of I and II, separately), which might be due to post-translational modification including the characteristic C-terminal cleavage at these two proteins as similar to that found in rat LC3 and yeast Atg8. Subcellular localization demonstrated that both LC3A and LC3B are colocalized with LC3 and associated with the autophagic membranes. Mutation analysis further revealed that the conserved Gly120 residues of LC3A and LC3B are essential for their characteristic C-terminal cleavage and localization to autophagic membranes. Present data suggested that LC3A and LC3B could also be used as two novel autophagosomal markers.

Original languageEnglish (US)
Pages (from-to)437-442
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume339
Issue number1
DOIs
StatePublished - Jan 6 2006

Keywords

  • Autophagy
  • LC3A
  • LC3B
  • Post-translational modification
  • Rat LC3
  • Subcellular localization

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Molecular cloning and characterization of rat LC3A and LC3B - Two novel markers of autophagosome'. Together they form a unique fingerprint.

  • Cite this