Isolation of cDNA clones encoding rat glial fibrillary acidic protein: expression in astrocytes and in Schwann cells.
Glial fibrillary acidic protein (GFAP) expressed by astrocytes in the central nervous system (CNS) has been extensively characterized but the molecular identity of related molecules in the peripheral nervous system (PNS) remains unclear. To examine possible structural differences between CNS and PNS GFAP, we have isolated cDNA clones for rat GFAP from both cultured astrocyte and Schwann cell libraries. Nucleotide sequence analysis indicated that the PNS and CNS GFAP clones contained identical coding regions, with a predicted protein product of 430 amino acids. However, the 5'-untranslated region of clone rGFA15, isolated from the Schwann cell library, was longer than that predicted for brain-derived GFAP mRNA. Primer extension analysis of RNA isolated from the RT4-D6 Schwann cell line indicated that the start site for PNS GFAP mRNA lies 169 bases upstream from that used in the CNS. In addition, tryptic peptide mapping of GFAP prepared from cultured astrocytes and Schwann cells revealed one major peptide fragment present in CNS GFAP but absent from PNS GFAP. These results suggest structural differences between GFAP in these two cell types, at both the nucleic acid and protein level, and are consistent with previous observations of immunochemical differences existing between CNS and PNS GFAP.
Feinstein
D L
DL
Department of Neuropharmacology, Scripps Research Institute, La Jolla, California.
Weinmaster
G A
GA
Milner
R J
RJ
D000595:Amino Acid Sequence; D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004247:DNA / Q000302:isolation & purification*; D015723:Gene Library; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics; D010449:Peptide Mapping; D012333:RNA, Messenger / Q000302:isolation & purification; D051381:Rats; D012583:Schwann Cells / Q000378:metabolism*; D014158:Transcription, Genetic
D005904:Glial Fibrillary Acidic Protein; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D004247:DNA