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Metabolic changes in the poly(ADP-ribosyl)ation pathway of differentiating rat germinal cells
- Source :
- Archives of biochemistry and biophysics. 381(1)
- Publication Year :
- 2000
-
Abstract
- Endogenous levels of poly(ADP-ribose) and betaNAD+ have been determined in rat male germinal cells at different stages of differentiation. The levels of both metabolites decreased progressively from primary spermatocytes to secondary spermatocytes and especially in spermatids. We have also determined the size and complexity of the ADP-ribose polymers synthesized in permeabilized germ cells. Polymers of different chain length and complexity were observed in cells incubated with different concentrations of [32P]betaNAD+; short polymers characterized primary spermatocytes incubated with low betaNAD+ concentration. In all cell fractions, polymers of over 20 residues in size were observed at high betaNAD+ levels. Long polymers were associated with the sulfuric acid-insoluble proteins (nonhistone proteins such as PARP itself). By contrast, oligomers of 20 ADP-ribose units or less were found in the sulfuric acid-soluble proteins (histone proteins). We have also identified the main ADP-ribose protein acceptors formed in each cell type. In all cells examined, PARP appears to be extensively automodified. However, by far, the H1t variant of histone H1 appeared to be the preferred ADP-ribose target among the acid-soluble proteins separated by reverse-phase HPLC. Therefore, we conclude that an active protein-poly(ADP-ribosyl)ation system is concentrated in primary spermatocytes, based on a high level of PARP automodification accompanied by the preferential heteromodification of the histone H1 variant specifically expressed in the cells undergoing the pachytene phase of the meiotic division.
- Subjects :
- Male
Cell type
Poly Adenosine Diphosphate Ribose
Cellular differentiation
Poly ADP ribose polymerase
Biophysics
Biology
In Vitro Techniques
Biochemistry
Histones
Non-histone protein
Histone H1
Spermatocytes
Animals
Rats, Wistar
Spermatogenesis
Molecular Biology
Proteins
Cell Differentiation
NAD
Spermatids
Spermatozoa
Rats
Histone
biology.protein
NAD+ kinase
Subjects
Details
- ISSN :
- 00039861
- Volume :
- 381
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Archives of biochemistry and biophysics
- Accession number :
- edsair.doi.dedup.....d407c6215227dd57e046aa408dd9592c