Structure of the gene encoding VGF, a nervous system-specific mRNA that is rapidly and selectively induced by nerve growth factor in PC12 cells.

Abstract

Nerve growth factor (NGF) plays a critical role in the development and survival of neurons in the peripheral nervous system. Following treatment with NGF but not epidermal growth factor, rat pheochromocytoma (PC12) cells undergo neural differentiation. We have cloned a nervous system-specific mRNA, NGF33.1, that is rapidly and relatively selectively induced by treatment of PC12 cells with NGF and basic fibroblast growth factor in comparison with epidermal growth factor. Analysis of the nucleic acid and predicted amino acid sequences of the NGF33.1 cDNA clone suggested that this clone corresponded to the NGF-inducible mRNA called VGF (A. Levi, J. D. Eldridge, and B. M. Paterson, Science 229:393-395, 1985; R. Possenti, J. D. Eldridge, B. M. Paterson, A. Grasso, and A. Levi, EMBO J. 8:2217-2223, 1989). We have used the NGF33.1 cDNA clone to isolate and characterize the VGF gene, and in this paper we report the complete sequence of the VGF gene, including 853 bases of 5' flank revealed TATAA and CCAAT elements, several GC boxes, and a consensus cyclic AMP response element-binding protein binding site. The VGF promoter contains sequences homologous to other NGF-inducible, neuronal promoters. We further show that VGF mRNA is induced in PC12 cells to a greater extent by depolarization and by phorbol-12-myristate-13-acetate treatment than by 8-bromo-cyclic AMP treatment. By Northern (RNA) and RNase protection analysis, VGF mRNA is detectable in embryonic and postnatal central and peripheral nervous tissues but not in a number of nonneural tissues. In the cascade of events which ultimately leads to the neural differentiation of NGF-treated PC12 cells, the VGF gene encodes the most rapidly and selectively regulated, nervous-system specific mRNA yet identified.

Degradable PRGD/PDLLA/β-TCP/NGF composites promote differentiation and regulate gene expression in rat pheochromocytoma cells

“…Surprisingly, however, the VGF mRNA in the NGF-embedded composite group showed a three-fold higher level of expression compared with the 50 ng/mL NGF group on day 2. Our results demonstrated that VGF mRNA levels were maximally induced to ~16-fold, which was in agreement with a previous report [22]. However, polarization can also lead to a rapid increase in VGF mRNA expression level by triggering calcium influx through voltage-gated calcium channels [22,23].…”

“…Our results demonstrated that VGF mRNA levels were maximally induced to ~16-fold, which was in agreement with a previous report [22]. However, polarization can also lead to a rapid increase in VGF mRNA expression level by triggering calcium influx through voltage-gated calcium channels [22,23]. During the degradation of the composites, β-TCP elevates the calcium concentration in the culture medium, which is involved in mediating the growth and migratory direction of axons.…”

Replication of linkage on chromosome 7q22 and association of the regional Reelin gene with working memory in schizophrenia families

“…82,83 We failed to show association to any of the candidate genes according to the dichotomized clinical diagnosis of schizophrenia and schizophrenia spectrum disorders among 245 schizophrenia families, originating from the same geographical region as the families revealing the linkage to 7q21-32. With the markers used, we had fairly good coverage of the allelic diversity of the four studied genes.…”

A Prohormone Convertase Cleavage Site within a Predicted α-Helix Mediates Sorting of the Neuronal and Endocrine Polypeptide VGF into the Regulated Secretory Pathway

“…A related series of constructs was made by truncating the full-length HA-tagged VGF using PCR primers to introduce termination codons following amino acids 177, 282, 367, 480, and 543 of rat VGF. The VGF:GFP constructs, which directly fuse VGF and GFP sequences in the vector pcDNA3 (Invitrogen) at the indicated sites, were created by PCR using pEGFP-N1 and the rat VGF cDNA (7,16) as templates. Sequences of all PCR-generated fragments were verified.…”

“…Secreted via the Regulated Release Pathway-VGF can be differentially processed, and the C terminus is rich in paired basic amino acids (7,9,16). Additionally, N-terminal signal peptide sequences and loop structures have been implicated in sorting, so two series of plasmids that contained either N-or C-terminal modifications were constructed.…”