Glyceraldehyde-3-phosphate dehydrogenase is a nonhistone protein and a possible activator of transcription in neurons.
A single-stranded DNA-binding protein of Mr 35,000 (35K protein) was isolated from calf cerebral cortex by affinity chromatography on immobilized double-stranded and single-stranded DNA. Its localization in the nuclear compartment was demonstrated by immunohistochemistry. Previous studies had uncovered a homologous nonhistone chromosomal protein in the nuclei of rat cerebral cortex neurons, cerebellar neurons, oligodendrocytes, and liver cells. The rat protein accumulated in the nuclear compartment of neurons in exact temporal coincidence with the arrest of cell division and the initiation of terminal differentiation. Therefore, in the present work, the 35K protein was tested for an activating role in RNA transcription. During the course of this study we became aware that the 35K protein was identical to a glycolytic enzyme, glyceraldehyde-3-phosphate dehydrogenase (GAPDH, EC 1.2.1.12). When authentic GAPDH from rabbit skeletal muscle was injected into Xenopus laevis oocytes, it greatly stimulated RNA polymerase II transcription, whereas the 35K protein from calf brain did not. This apparent discrepancy was partially resolved by the finding that rabbit muscle GAPDH could be fractionated into two components by affinity chromatography on single-stranded DNA cellulose. Only 5% of the applied protein was retained on the column and could be eluted with a shallow salt gradient identical to the one used for the isolation of the 35K protein. This single-stranded DNA-binding component of rabbit muscle GAPDH did not stimulate transcription. Apparently, the 35K protein from calf brain corresponded to this single-stranded DNA-binding subfraction, which explained its failure to activate transcription.(ABSTRACT TRUNCATED AT 250 WORDS)
G
G
Morgenegg
G C
GC
Winkler
U
U
Hübscher
C W
CW
Heizmann
J
J
Mous
C C
CC
Kuenzle
D000818:Animals; D002417:Cattle; D002467:Cell Nucleus / Q000201:enzymology; D002540:Cerebral Cortex / Q000201:enzymology; D055598:Chemical Phenomena; D002621:Chemistry; D006224:Cricetinae; D003412:Cricetulus; D003593:Cytoplasm / Q000201:enzymology; D004261:DNA Replication / Q000187:drug effects; D004277:DNA, Single-Stranded / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000302:isolation & purification / Q000494:pharmacology / Q000502:physiology*; D009132:Muscles / Q000201:enzymology; D009474:Neurons / Q000502:physiology*; D009836:Oligodendroglia / Q000201:enzymology / Q000648:ultrastructure; D009865:Oocytes / Q000187:drug effects; D012313:RNA / Q000235:genetics; D011817:Rabbits; D051381:Rats; D013268:Stimulation, Chemical; D014158:Transcription, Genetic / Q000187:drug effects; D014982:Xenopus laevis
D004277:DNA, Single-Stranded; D012313:RNA; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases