Dystonia type 6 gene product Thap1: identification of a 50 kDa DNA-binding species in neuronal nuclear fractions.

Acta Neuropathol Commun 2014 Sep 18;2:139. Epub 2014 Sep 18.

Mutations in THAP1 result in dystonia type 6, with partial penetrance and variable phenotype. The goal of this study was to examine the nature and expression pattern of the protein product(s) of the Thap1 transcription factor (DYT6 gene) in mouse neurons, and to study the regional and developmental distribution, and subcellular localization of Thap1 protein. The goal was accomplished via overexpression and knock-down of Thap1 in the HEK293T cell line and in mouse striatal primary cultures and western blotting of embryonic Thap1-null tissue. The endogenous and transduced Thap1 isoforms were characterized using three different commercially available anti-Thap1 antibodies and validated by immunoprecipitation and DNA oligonucleotide affinity chromatography. We identified multiple, novel Thap1 species of apparent Mr 32 kDa, 47 kDa, and 50-52 kDa in vitro and in vivo, and verified the previously identified species at 29-30 kDa in neurons. The Thap1 species at the 50 kDa size range was exclusively detected in murine brain and testes and were located in the nuclear compartment. Thus, in addition to the predicted 25 kDa apparent Mr, we identified Thap1 species with greater apparent Mr that we speculate may be a result of posttranslational modifications. The neural localization of the 50 kDa species and its nuclear compartmentalization suggests that these may be key Thap1 species controlling neuronal gene transcription. Dysfunction of the neuronal 50 kDa species may therefore be implicated in the pathogenesis of DYT6.

Download full-text PDF

Source
http://dx.doi.org/10.1186/s40478-014-0139-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4177242PMC
September 2014
14 Reads

Publication Analysis

Top Keywords

thap1 species
16
thap1
10
species
8
50 kda species
8
dystonia type
8
greater apparent
4
endogenous transduced
4
tissue endogenous
4
thap1-null tissue
4
transduced thap1
4
embryonic thap1-null
4
apparent speculate
4
isoforms characterized
4
apparent identified
4
commercially anti-thap1
4
25 kda apparent
4
three commercially
4
identified thap1
4
thap1 isoforms
4
speculate result
4

References

(Supplied by CrossRef)

SB Bressman et al.
Lancet Neurol 2009

A Djarmati et al.
Lancet Neurol 2009

T Fuchs et al.
Nat Genet 2009

MS LeDoux et al.
Parkinsonism Relat Disord 2012

G Xiromerisiou et al.
Mov Disord 2012

LP Deiss et al.
Genes Dev 1995

M Gale Jr et al.
Mol Cell Biol 1998

Y Lin et al.
J Biol Chem 2002

M Roussigne et al.
Oncogene 2003

M Roussigne et al.
Trends Biochem Sci 2003

T Clouaire et al.
Proc Natl Acad Sci U S A 2005

Similar Publications