原创力文档a sequence-dependent exonuclease activity from tetrahymena thermophila一个顺序相依核酸外切酶活性从四膜虫thermophila.pdfVIP
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a sequence-dependent exonuclease activity from tetrahymena thermophila一个顺序相依核酸外切酶活性从四膜虫thermophila
Tom and Greider BMC Biochemistry 2010, 11:45
/1471-2091/11/45
RESEARCH ARTICLE Open Access
A sequence-dependent exonuclease activity from
Tetrahymena thermophila
*
Hui-I Kao Tom, Carol W Greider
Abstract
Background: Telomere function requires a highly conserved G rich 3’- overhang. This structure is formed by 5’-
resection of the C-rich telomere strand. However, while many nucleases have been suggested to play a role in
processing, it is not yet clear which nucleases carry out this 5’-resection.
Results: We used biochemical purification to identify a sequence-dependent exonuclease activity in Tetrahymena
thermophila cell extracts. The nuclease activity showed specificity for 5’-ends containing AA or AC sequences, unlike
Exo1, which showed sequence-independent cleavage. The Tetrahymena nuclease was active on both
phosphorylated and unphosphorylated substrates whereas Exo1 requires a 5’-phosphate for cleavage.
Conclusions: The specificities of the enzyme indicate that this novel Tetrahymena exonuclease is distinct from
Exo1 and has properties required for 3’-overhang formations at telomeres.
Background mammals and other species. Loss of the overhangs due
Functional telomeres are essential for cell survival. Telo- to disruption of the telomere complex, termed shelterin,
meres distinguish natural chromosome ends from DNA leads to telomere-induced DNA damage response and
breaks. When telomere function is lost, cells arrest and chromosome end-to-end fusion [22,23]. In addition to
undergo senescence or apoptosis [1-6]. Normal DNA maintenance of DNA ends, the 3’-overhang is essential
replication causes telomere shortening, which is coun-
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