The yeast STM1 gene encodes a purine motif triple helical DNA-binding protein.
The formation of triple helical DNA has been evoked in several cellular processes including transcription, replication, and recombination. Using conventional and affinity chromatography, we purified from Saccharomyces cerevisiae whole-cell extract a 35-kDa protein that avidly and specifically bound a purine motif triplex (with a K(d) of 61 pM) but not a pyrimidine motif triplex or duplex DNA. Peptide microsequencing identified this protein as the product of the STM1 gene. Confirmation that Stm1p is a purine motif triplex-binding protein was obtained by electrophoretic mobility shift assays using either bacterially expressed, recombinant Stm1p or whole-cell extracts from stm1Delta yeast. Stm1p has previously been identified as G4p2, a G-quartet nucleic acid-binding protein. This suggests that some proteins actually recognize features shared by G4 DNA and purine motif triplexes, e.g. Hoogsteen hydrogen-bonded guanines. Genetically, the STM1 gene has been identified as a multicopy suppressor of mutations in several genes involved in mitosis (e.g. TOM1, MPT5, and POP2). A possible role for multiplex DNA and its binding proteins in mitosis is discussed.
Nelson
L D
LD
Department of Cancer Biology, The University of Texas M.D. Anderson Cancer, Houston, Texas 77030, USA.
Musso
M
M
Van Dyke
M W
MW
D001120:Arginine / Q000378:metabolism; D001667:Binding, Competitive; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D039642:Eukaryotic Initiation Factors; D005656:Fungal Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology; D007700:Kinetics; D008239:Lysine / Q000378:metabolism; D008957:Models, Genetic; D010448:Peptide Initiation Factors; D011687:Purines / Q000378:metabolism; D012316:RNA Nucleotidyltransferases / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins
D004268:DNA-Binding Proteins; D039642:Eukaryotic Initiation Factors; D005656:Fungal Proteins; D010448:Peptide Initiation Factors; D011687:Purines; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; C083432:TIF3 protein, S cerevisiae; C051802:triplex DNA; D004247:DNA; D001120:Arginine; D012316:RNA Nucleotidyltransferases; D008239:Lysine