PMID: 2118523Sep 5, 1990Paper

The zinc finger protein NGFI-A exists in both nuclear and cytoplasmic forms in nerve growth factor-stimulated PC12 cells.

The Journal of Biological Chemistry
M L DayJ Milbrandt

Abstract

The NGFI-A gene, which encodes a zinc finger protein with a predicted molecular mass of congruent to 54 kDa, is rapidly activated in PC12 cells by nerve growth factor (NGF). As a transcription factor, NGFI-A is a potentially important mediator of the cellular response to this growth factor. To characterize this protein, antibodies to four different domains of NGFI-A were raised. Immunogens included a bacterial trpE/NGFI-A fusion protein (A310) and three peptides predicted from the NGFI-A cDNA nucleotide sequence. Three of these antisera recognize two predominant NGFI-A species in NGF-stimulated PC12 cells: a 54-kDa form and a closely spaced doublet at 84 kDa. However, one of these antisera, directed against the last 15 carboxyl terminal residues of NGFI-A, recognizes only the 84-kDa species. Both NGFI-A species are rapidly induced in PC12 cells by NGF, phorbol ester, and the calcium ionophore A23187. Pulse-chase analysis revealed no obvious precursor-product relationship between these two forms and demonstrated that both the 54- and 84-kDa species are short-lived proteins. V8 protease digestion of the 54- and 84-kDa forms resulted in the formation of several small peptides that were common to both species. The digest of each sp...Continue Reading

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