Abnormal interaction of motor neuropathy-associated mutant HspB8 (Hsp22) forms with the RNA helicase Ddx20 (gemin3).

Abstract

A number of missense mutations in the two related small heat shock proteins HspB8 (Hsp22) and HspB1 (Hsp27) have been associated with the inherited motor neuron diseases (MND) distal hereditary motor neuropathy and Charcot-Marie-Tooth disease. HspB8 and HspB1 interact with each other, suggesting that these two etiologic factors may act through a common biochemical mechanism. However, their role in neuron biology and in MND is not understood. In a yeast two-hybrid screen, we identified the DEAD box protein Ddx20 (gemin3, DP103) as interacting partner of HspB8. Using co-immunoprecipitation, chemical cross-linking, and in vivo quantitative fluorescence resonance energy transfer, we confirmed this interaction. We also show that the two disease-associated mutant HspB8 forms have abnormally increased binding to Ddx20. Ddx20 itself binds to the survival-of-motor-neurons protein (SMN protein), and mutations in the SMN1 gene cause spinal muscular atrophy, another MND and one of the most prevalent genetic causes of infant mortality. Thus, these protein interaction data have linked the three etiologic factors HspB8, HspB1, and SMN protein, and mutations in any of their genes cause the various forms of MND. Ddx20 and SMN protein are involved in spliceosome assembly and pre-mRNA processing. RNase treatment affected the interaction of the mutant HspB8 with Ddx20 suggesting RNA involvement in this interaction and a potential role of HspB8 in ribonucleoprotein processing.

• Authors: Xiankui Sun, Jean-Marc Fontaine, Adam D Hoppe, Serena Carra, Cheryl DeGuzman, Jody L Martin, Stephanie Simon, Patrick Vicart, Michael J Welsh, Jacques Landry, Rainer Benndorf
• Journal: Cell stress & chaperones (Cell Stress Chaperones) Vol. 15 Issue 5 Pg. 567-82 (Sep 2010) ISSN: 1466-1268 [Electronic] Netherlands
• PMID: 20157854 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• HSP27 Heat-Shock Proteins
• HSPB1 protein, human
• HSPB8 protein, human
• Heat-Shock Proteins
• Molecular Chaperones
• Survival of Motor Neuron 1 Protein
• Protein Serine-Threonine Kinases
• DDX20 protein, human
• DEAD Box Protein 20

Topics

• Amino Acid Sequence
• Cell Line
• Charcot-Marie-Tooth Disease (metabolism)
• DEAD Box Protein 20 (chemistry, genetics, metabolism)
• Fluorescence Resonance Energy Transfer
• Fluorescent Antibody Technique
• HSP27 Heat-Shock Proteins (chemistry, genetics, metabolism)
• Heat-Shock Proteins (chemistry, genetics, metabolism)
• Humans
• Immunoprecipitation
• Isoelectric Focusing
• Molecular Chaperones
• Molecular Sequence Data
• Protein Serine-Threonine Kinases (chemistry, genetics, metabolism)
• Survival of Motor Neuron 1 Protein (chemistry, genetics, metabolism)
• Two-Hybrid System Techniques