Detection of telomerase activity in tissues and primary cultured lymphoid cells of Penaeus japonicus.
Telomerase is a ribonucleoprotein enzyme that can elongate telomeric DNA, which is thought to be required for the development of cellular immortality and oncogenesis in mammals. We examined telomerase activity in tissues and primary cultured lymphoid cells of adult penaeid shrimps. Using the telomeric repeat amplification protocol (TRAP), we studied the characteristics of a putative novel telomerase in Penaeus japonicus. This telomerase could be inactivated by heating or treatment with RNase A or proteinase K. At elongation, this telomerase required dATP, dGTP, and dTTP, but not dCTP, as substrates. Sequence analysis of the TRAP product revealed that this telomerase synthesized (TTAGG)(n) repeated sequences. The activity of this telomerase was decreased but still readily detectable in 100 ng of protein extract from lymphoid tissue. The telomerase activity was detected in all examined tissues including testis, ovary, lymphoid, heart, hepatopancreas, and muscle. The highest telomerase activity was in the extract of ovarian tissues. In primary cultured lymphoid cells, the telomerase activity was retained. Thus, primary cultured lymphoid cells of Penaeus japonicus possess one of the factors necessary for cell line establishment.
Lang
Gang-Hua
GH
Institute of Applied Biochemistry, University of Tsukuba, Tennodai 1-1-1, Tsukuba, Ibaraki 305-8572, Japan.
Wang
Yong
Y
Nomura
Nakao
N
Matsumura
Masatoshi
M
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D019286:Endopeptidase K / Q000378:metabolism; D008214:Lymphocytes / Q000201:enzymology*; D021141:Nucleic Acid Amplification Techniques; D033561:Penaeidae / Q000201:enzymology*; D012259:Ribonuclease, Pancreatic / Q000378:metabolism; D017422:Sequence Analysis, DNA; D019098:Telomerase / Q000378:metabolism*; D016615:Telomere / Q000235:genetics; D013997:Time Factors
D017931:DNA Primers; D019098:Telomerase; D012259:Ribonuclease, Pancreatic; D019286:Endopeptidase K