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Reverse transcriptase activity from human embryonal carcinoma cells NTera2D1.
Author(s) -
Deragon J. M.,
Sinnett D.,
Labuda D.
Publication year - 1990
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1990.tb07537.x
Subject(s) - service (business) , library science , humanities , art , computer science , economy , economics
We have identified an RNA‐dependent DNA polymerase activity in the microsomal fraction of human pluri‐potential embryonal carcinoma cells NTera2D1, which are known to express the full length coding strand of the genomic Line‐1 (L1) elements. This activity was classified as a reverse transcriptase (RT) based on its utilization of an RT specific synthetic poly(Cm) template in the presence of Mn2+ ions. Treatment of the cell by ultraviolet irradiation (200 erg/mm2) which resulted in a 2‐ to 3‐fold enhancement of the RT activity, was required for the reproducible detection of the activity throughout the entire purification procedure. More than a 100‐fold enrichment in RT activity was obtained by centrifugation in a glycerol step gradient and a linear sucrose density gradient followed by Sephacryl S‐1000 gel filtration. These experiments demonstrated that the RT activity was associated with a macromolecular complex having the characteristics of a viral‐like particle with a major protein component of 37 kd. The presence of L1 mRNA in RT‐containing fractions suggests that the activity identified could originate from L1 elements and/or be involved in the mechanism of retroposition.