Heterogeneity of the high molecular weight tau proteins in N115 neuroblastoma cells.
The sequence of a high molecular weight (HMW) tau cDNA cloned from a neuroblastoma N115 library contains, in addition to the C- and N-terminal and middle regions present in the low molecular weight mouse brain tau proteins, a 711-bp nonhomologous domain (exon 4a) and a region of 198 bp corresponding to exon 6 of the tau gene. Protein immunoblot analysis, performed with antibodies specific either for a sequence present in the N-terminal region of all the tau variants or for exon 4a revealed several bands suggesting that more than one tau form is expressed in this cell line. Northern blot experiments performed with a number of cDNA probes spanning domains common and uncommon to low molecular weight and HMW tau allowed the identification of four tau transcripts differing in the size of their coding and noncoding regions. All these transcripts contain the sequence encoded by exon 6, but two of them lack exon 4a. As shown by RNase protection assays, the N-terminal region of these transcripts is also variable and contains either exon 1, or exons 1 and 2, or exons 1-3. Yet all these HMW tau forms contain four homologous repeats in their C-terminal domain both in the differentiated and nondifferentiated cells, i.e., have adult characteristics. In conclusion, the data reported in this article demonstrate that several HMW tau variants are expressed in neuroblastoma N115 cells and that the transition between immature to mature tau forms occurring during brain development is not required for neurite outgrowth during morphological differentiation of this cell line.
Gache
Y
Y
INSERM U 282-CNRS, Hôpital Henri Mondor, Créteil, France.
Guilleminot
J
J
Bridoux
A M
AM
Nunez
J
J
D000818:Animals; D001483:Base Sequence; D015151:Immunoblotting; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009447:Neuroblastoma / Q000378:metabolism* / Q000473:pathology; D016133:Polymerase Chain Reaction; D012260:Ribonucleases; D014407:Tumor Cells, Cultured; D016875:tau Proteins / Q000737:chemistry* / Q000378:metabolism*
D015335:Molecular Probes; D016875:tau Proteins; D012260:Ribonucleases