Reconstitution of telomerase activity utilizing human catalytic subunit expressed in insect cells.
Telomerase is a specialized reverse transcriptase responsible for maintaining the termini of linear chromosomes. The human enzyme is a ribonucleoprotein complex minimally comprising a catalytic protein moiety (hTERT) and an RNA subunit (hTR) which acts as the template for the reverse transcriptase reaction. Here we report expression of recombinant hTERT protein in insect cells utilizing a baculovirus expression system. The recombinant hTERT protein reconstitutes telomerase activity in the presence of hTR, either when co-expressed in insect cells or when added in vitro. Reconstitution of telomerase activity using this system will facilitate further analysis of the biochemical and biophysical properties of this enzyme.
Mikuni
Osamu
O
Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, 194-8533, Tokyo, Japan. osamu.mikuni@kyowa.co.jp
Trager
James B
JB
Ackerly
Heidi
H
Weinrich
Scott L
SL
Asai
Akira
A
Yamashita
Yoshinori
Y
Mizukami
Tamio
T
Anazawa
Hideharu
H
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D002384:Catalysis; D002460:Cell Line; D004268:DNA-Binding Proteins; D004789:Enzyme Activation; D006801:Humans; D021122:Protein Subunits; D011994:Recombinant Proteins / Q000378:metabolism; D012995:Solubility; D018411:Spodoptera / Q000235:genetics*; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism*
D004268:DNA-Binding Proteins; D021122:Protein Subunits; D011994:Recombinant Proteins; D019098:Telomerase