This proposal will continue our studies on the structure and function of the neuronal intermediate filaments. Although our focus had been primarily on the neurofilament triplet proteins, NF-L, NF-M and NF-H, the last grant period has also yielded a wealth of data on alpha-internexin, another neuronal intermediate filament protein, first described in our laboratory. Over this time, we obtained cDNA and genomic clones, as well as monoclonal and polyclonal antibodies, and studied the in vivo and in vitro assembly of these neuronal intermediate filament proteins. We have shown that alpha-internexin is the first neuronal intermediate filament protein to be expressed by most if not all neurons during development. The first specific aim is to compare the importance of different structural domains of the neuronal intermediate filament proteins in in vivo and in vitro assembly. These studies will enable us to understand the mechanisms by which intermediate filaments are formed in neurons and should begin to explain the importance of the diversity of neuronal intermediate filaments.
The second aim i s to determine the control elements responsible for the tissue-specific expression of alpha- internexin.
This aim will allow us to determine how this neuron specific gene is regulated. It could also provide us with the tools necessary to target proteins specifically to the nervous system.
The third aim i s to accomplish the targeted disruption of the gene for alpha-internexin in transgenic mice. these experiments should help us in determining the function of this protein. The roles that neuronal intermediate filaments play in development, axonal integrity and neurological diseases are not clear, although they are implicated in all of them. Recent studies on the genetic skin diseases, Epidermolysis Bullosa Simplex and Epidermolytic Hyperkeratosis point to non-life threatening, but still debilitating illnesses resulting from mutations in the keratins. Similarly, neuronal intermediate filaments may not be essential for survival per se, but defects in neuronal intermediate filaments are likely to be related to neuronal diseases. In this proposal, we hope to determine the function of one of the neuronal intermediate filament proteins, alpha-internexin and identify its role in the nervous system and in neurological diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS015182-19
Application #
2445702
Study Section
Special Emphasis Panel (ZRG1-NEUB-1 (03))
Program Officer
Streicher, Eugene
Project Start
1987-07-01
Project End
1998-06-30
Budget Start
1997-07-01
Budget End
1998-06-30
Support Year
19
Fiscal Year
1997
Total Cost
Indirect Cost
Name
Columbia University (N.Y.)
Department
Pathology
Type
Schools of Medicine
DUNS #
167204994
City
New York
State
NY
Country
United States
Zip Code
10032
Liem, Ronald K H; Messing, Albee (2009) Dysfunctions of neuronal and glial intermediate filaments in disease. J Clin Invest 119:1814-24
Ching, Gee Y; Liem, Ronald K H (2009) RE1 silencing transcription factor is involved in regulating neuron-specific expression of alpha-internexin and neurofilament genes. J Neurochem 109:1610-23
Goryunov, Dmitry; Nightingale, Andrew; Bornfleth, Lorelei et al. (2008) Multiple disease-linked myotubularin mutations cause NFL assembly defects in cultured cells and disrupt myotubularin dimerization. J Neurochem 104:1536-52
Kabzinska, Dagmara; Perez-Olle, Raul; Goryunov, Dmitry et al. (2006) Is a novel I214M substitution in the NEFL gene a cause of Charcot-Marie-Tooth disease? Functional analysis using cell culture models. J Peripher Nerv Syst 11:225-31
Perez-Olle, Raul; Lopez-Toledano, Miguel A; Goryunov, Dmitry et al. (2005) Mutations in the neurofilament light gene linked to Charcot-Marie-Tooth disease cause defects in transport. J Neurochem 93:861-74
Perez-Olle, Raul; Jones, Sidonie T; Liem, Ronald K H (2004) Phenotypic analysis of neurofilament light gene mutations linked to Charcot-Marie-Tooth disease in cell culture models. Hum Mol Genet 13:2207-20
Perez-Olle, Raul; Lopez-Toledano, Miguel A; Liem, Ronald K H (2004) The G336S variant in the human neurofilament-M gene does not affect its assembly or distribution: importance of the functional analysis of neurofilament variants. J Neuropathol Exp Neurol 63:759-74
Leung, Conrad L; He, Cui Zhen; Kaufmann, Petra et al. (2004) A pathogenic peripherin gene mutation in a patient with amyotrophic lateral sclerosis. Brain Pathol 14:290-6
Leung, Conrad L; Green, Kathleen J; Liem, Ronald K H (2002) Plakins: a family of versatile cytolinker proteins. Trends Cell Biol 12:37-45
Perez-Olle, Raul; Leung, Conrad L; Liem, Ronald K H (2002) Effects of Charcot-Marie-Tooth-linked mutations of the neurofilament light subunit on intermediate filament formation. J Cell Sci 115:4937-46

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