Purification and characterization of Stn1p, a single-stranded telomeric DNA binding protein.
In Saccharomyces cerevisiae, Stn1p and Ten1p are required for telomere maintenance. These two proteins and another telomeric single-stranded DNA binding protein, Cdc13p, have been proposed to form a complex to control telomere integrity. In this work, we purified the recombinant Stn1p in Escherichia coli and found that the purified protein could specifically interact with single-stranded telomeric DNA in vitro. Co-fractionation of co-overexpressed Stn1p and Ten1p in insect cells revealed their stable association. A Stn1p/Ten1p binary complex was reconstituted with purified recombinant proteins in vitro. These results indicated that Stn1p and Ten1p interact with each other directly, which is important in telomere length regulation and end protection.
Qian
Wei
W
Department of Biochemistry, School of Medicine, Nantong University, Nantong, Jiangsu 226001, China. weiqianhuang@hotmail.com
Fu
Xiao-Hong
XH
Zhou
Jin-Qiu
JQ
D000818:Animals; D001483:Base Sequence; D018797:Cell Cycle Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D016615:Telomere / Q000235:genetics / Q000378:metabolism*; D034501:Telomere-Binding Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*
D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D004277:DNA, Single-Stranded; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C428109:Stn1 protein, S cerevisiae; C512860:TEN1 protein, S cerevisiae; D034501:Telomere-Binding Proteins