TY - JOUR
T1 - The modified base J is the target for a novel DNA-binding protein in kinetoplastid protozoans
AU - Cross, Mike
AU - Kieft, Rudo
AU - Sabatini, Robert
AU - Wilm, Matthias
AU - de Kort, Martin
AU - van der Marel, Gijs A.
AU - van Boom, Jacques H.
AU - van Leeuwen, Fred
AU - Borst, Piet
PY - 1999
Y1 - 1999
N2 - DNA from Kinetoplastida contains the unusual modified base β-D-glucosyl(hydroxymethyl))uracil, called J. Base J is found predominantly in repetitive DNA and correlates with epigenetic silencing of telomeric variant surface glycoprotein genes in Trypanosoma brucei. We have now identified a protein in nuclear extracts of bloodstream stage T. brucei that binds specifically to J-containing duplex DNA. J-specific DNA binding was also observed with extracts from the kinetoplastids Crithidia fasciculata and Leishmania tarentolae. We purified the 90 kDa C. fasciculata J-binding protein 50,000-fold and cloned the corresponding gene from C. fasciculata, T. brucei and L. tarentolae. Recombinant proteins expressed in Escherichia coli demonstrated J-specific DNA binding. The J-binding proteins show 43-63% identity and are unlike any known protein. The discovery of a J-binding protein suggests that J, like methylated cytosine in higher eukaryotes, functions via a protein intermediate.
AB - DNA from Kinetoplastida contains the unusual modified base β-D-glucosyl(hydroxymethyl))uracil, called J. Base J is found predominantly in repetitive DNA and correlates with epigenetic silencing of telomeric variant surface glycoprotein genes in Trypanosoma brucei. We have now identified a protein in nuclear extracts of bloodstream stage T. brucei that binds specifically to J-containing duplex DNA. J-specific DNA binding was also observed with extracts from the kinetoplastids Crithidia fasciculata and Leishmania tarentolae. We purified the 90 kDa C. fasciculata J-binding protein 50,000-fold and cloned the corresponding gene from C. fasciculata, T. brucei and L. tarentolae. Recombinant proteins expressed in Escherichia coli demonstrated J-specific DNA binding. The J-binding proteins show 43-63% identity and are unlike any known protein. The discovery of a J-binding protein suggests that J, like methylated cytosine in higher eukaryotes, functions via a protein intermediate.
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=0033570918&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/10562569
U2 - https://doi.org/10.1093/emboj/18.22.6573
DO - https://doi.org/10.1093/emboj/18.22.6573
M3 - Article
C2 - 10562569
SN - 0261-4189
VL - 18
SP - 6573
EP - 6581
JO - EMBO Journal
JF - EMBO Journal
IS - 22
ER -