MAP4 mechanism that stabilizes mitochondrial permeability transition in hypoxia: microtubule enhancement and DYNLT1 interaction with VDAC1.

Abstract

Mitochondrial membrane permeability has received considerable attention recently because of its key role in apoptosis and necrosis induced by physiological events such as hypoxia. The manner in which mitochondria interact with other molecules to regulate mitochondrial permeability and cell destiny remains elusive. Previously we verified that hypoxia-induced phosphorylation of microtubule-associated protein 4 (MAP4) could lead to microtubules (MTs) disruption. In this study, we established the hypoxic (1% O(2)) cell models of rat cardiomyocytes, H9c2 and HeLa cells to further test MAP4 function. We demonstrated that increase in the pool of MAP4 could promote the stabilization of MT networks by increasing the synthesis and polymerization of tubulin in hypoxia. Results showed MAP4 overexpression could enhance cell viability and ATP content under hypoxic conditions. Subsequently we employed a yeast two-hybrid system to tag a protein interacting with mitochondria, dynein light chain Tctex-type 1 (DYNLT1), by hVDAC1 bait. We confirmed that DYNLT1 had protein-protein interactions with voltage-dependent anion channel 1 (VDAC1) using co-immunoprecipitation; and immunofluorescence technique showed that DYNLT1 was closely associated with MTs and VDAC1. Furthermore, DYNLT1 interactions with MAP4 were explored using a knockdown technique. We thus propose two possible mechanisms triggered by MAP4: (1) stabilization of MT networks, (2) DYNLT1 modulation, which is connected with VDAC1, and inhibition of hypoxia-induced mitochondrial permeabilization.

• Authors: Ya-dong Fang, Xue Xu, Yong-ming Dang, Yi-ming Zhang, Jia-ping Zhang, Jiong-yu Hu, Qiong Zhang, Xia Dai, Miao Teng, Dong-xia Zhang, Yue-sheng Huang
• Journal: PloS one (PLoS One) Vol. 6 Issue 12 Pg. e28052 ( 2011) ISSN: 1932-6203 [Electronic] United States
• PMID: 22164227 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• DYNLT1 protein, human
• Microtubule-Associated Proteins
• Recombinant Proteins
• VDAC1 protein, human
• Green Fluorescent Proteins
• Adenosine Triphosphate
• Voltage-Dependent Anion Channel 1
• Dyneins

Topics

• Adenosine Triphosphate (metabolism)
• Animals
• Cell Survival
• Dyneins (metabolism)
• Gene Expression Regulation
• Green Fluorescent Proteins (metabolism)
• HeLa Cells
• Humans
• Hypoxia (metabolism)
• Microscopy, Confocal (methods)
• Microtubule-Associated Proteins (metabolism)
• Microtubules (metabolism)
• Mitochondria (metabolism)
• Permeability
• Rats
• Rats, Sprague-Dawley
• Recombinant Proteins (metabolism)
• Two-Hybrid System Techniques
• Voltage-Dependent Anion Channel 1 (metabolism)