Publication: Intracellular distribution of the ΔNp73 protein isoform in medulloblastoma cells: a study with newly generated rabbit polyclonal antibodies
Authors
Veselska, Renata ; Neradil, Jakub ; Nekulova, Marta ; Dobrucka, Lucia ; Vojtesek, Borivoj ; Sterba, Jaroslav ; Zitterbart, Karel
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Publisher
F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología
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DOI
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info:eu-repo/semantics/article
Description
Abstract
The p73 protein is a member of the p53
family of transcription factors that has two N-terminal
isoforms: the TAp73 isoform is reported to have a tumor
suppressor function, whereas the ΔNp73 isoform likely
has oncogenic potential. The expression of these
isoforms and the differences in their intracellular
distribution have been described in many cancer types;
however, little is known about the p73 isoforms in brain
tumors. Our study is focused on the intracellular
localization of ΔNp73 in medulloblastoma cell lines.
Due to a lack of suitable anti-ΔNp73 antibodies, we
developed two new rabbit polyclonal antibodies, ΔNp73-
26 and Δ Np73-27, with sufficient specificity, as
demonstrated by immunodetection methods using
transiently transfected cell lines. Both of these new
antibodies were subsequently used for analysis of the
ΔNp73 distribution in medulloblastoma cells using
immunofluorescence, immunoblotting and immunogold
labeling for transmission electron microscopy. We found
a nuclear localization of the ΔNp73 isoform in all of the
medulloblastoma cell lines included in this study.
Furthermore, a non-random accumulation of the ΔNp73
isoform near the cell nuclei was observable in all of
these cell lines. By double-labeling with ΔNp73 and
golgin-97, we showed the co-localization of the ΔNp73
isoform with the Golgi apparatus. Nevertheless, further
detailed analyses of possible interactions of ΔNp73 with
the proteins accumulated in the Golgi apparatus should
be performed to explain the dynamics of ΔNp73 outside
the cell nucleus.
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