Nuclear matrix bound terminal deoxynucleotidyl transferase in rat thymus nuclei. I. A possible site for TdT mediated function.
Approximately 80% of the terminal deoxynucleotidyl transferase (TdT) in thymus glands from 3-4 week old rats was found to be localized in the nucleus and the remaining 20% in the cytosol. Following endogenous nuclease digestion of the thymus nuclei, 70-85% of the nuclear TdT could be removed by low salt and high salt extractions, whereas 15-30% of the enzyme remained tightly bound to the residual nuclear matrix. Low salt and high salt extracts of the nuclei contained a mixture of 58, 56, 45 and 44 kDa species of TdT whereas only 58 kDa species of the enzyme was found to be associated with the matrix. In addition to TdT, 20-25% of the nuclear DNA polymerase alpha was also tightly bound to the isolated nuclear matrix. These observations lead us to propose that besides being the site of DNA replication via-matrix bound replicational complexes [Van der Velden H.M.W. & Wanka F., Molecular Biology Reports 12 (1987): 69], nuclear matrix may also be the site of TdT mediated function and that matrix bound TdT and free TdT could be the functional and nonfunctional forms of the enzyme, respectively, in the thymus gland.
Pandey
V N
VN
Biochemistry Division, Bhabha Atomic Research Centre, Bombay, India.
Dave
V P
VP
Patil
M S
MS
D000345:Affinity Labels; D000818:Animals; D003600:Cytosol / Q000201:enzymology; D004247:DNA / Q000096:biosynthesis; D004253:DNA Nucleotidylexotransferase / Q000032:analysis*; D004259:DNA-Directed DNA Polymerase / Q000032:analysis; D066298:In Vitro Techniques; D007700:Kinetics; D015530:Nuclear Matrix / Q000201:enzymology*; D011233:Precipitin Tests; D011487:Protein Conformation; D051381:Rats; D011919:Rats, Inbred Strains; D013950:Thymus Gland / Q000201:enzymology*
D000345:Affinity Labels; D004247:DNA; D004253:DNA Nucleotidylexotransferase; D004259:DNA-Directed DNA Polymerase