Characterization of nucleoprotein complexes in plants with human-type telomere motifs.
A conserved feature of telomeres is the 3'-overhang of their G-rich strand. These G-overhangs function as substrates for telomerase-mediated strand extension, and are critical for end-protection of telomeres. These functions and their regulations are mediated by specific G-overhang binding proteins. In species of the plant order Asparagales, telomere motifs have diverged from a type typical of the plant Arabidopsis thaliana (TTTAGGG)(n) to a type typical of human (TTAGGG)(n). Presumably, this change in motif had an impact on the structure of the telomere and/or the binding of telomeric proteins, including the G-overhang binding proteins. Therefore, we analyse here nucleoprotein complexes formed by protein extracts from plants possessing human-type telomeres (Muscari armeniacum and Scilla peruviana). Proteins were characterized that bind to the G-rich strand of both telomere motifs, or to the ancestral Arabidopsis-type motif alone, but none bound to double-stranded or C-rich complementary strand telomere motifs. We demonstrate the size, sequence-specificity and thermostability of these DNA-binding proteins. We also analysed the formation of complexes from renatured protein fractions after SDS-PAGE (sodium-dodecyl-sulphate polyacrylamide-gel-electrophoresis). We discuss the evolutionary consequences of protein binding flexibility, to act on both ancestral and present telomeric sequences. Of particular interest is that the ancestral repeat, which is thought not to form the telomere, binds the proteins most strongly. These data are discussed in line with other known plant telomere-binding proteins and with the complex nature of the telomere in Asparagales carrying a human-type motif.
Rotková
Gabriela
G
Institute of Biophysics, Královopolská 135, CZ-61265 Brno, Czech Republic.
Sýkorová
Eva
E
Fajkus
Jirí
J
D001483:Base Sequence; D018744:DNA, Plant / Q000235:genetics; D006801:Humans; D019668:Liliaceae / Q000235:genetics*; D009698:Nucleoproteins / Q000235:genetics*; D010940:Plant Proteins; D010944:Plants; D011489:Protein Denaturation; D016615:Telomere
D018744:DNA, Plant; D009698:Nucleoproteins; D010940:Plant Proteins