Plant chromosomal HMGB proteins efficiently promote the bacterial site-specific beta-mediated recombination in vitro and in vivo.
In the presence of an accessory DNA bending protein, the bacterial site-specific beta recombinase catalyzes resolution and DNA inversion. Five different maize high mobility group B (HMGB) proteins were examined for their potential to facilitate beta recombination in vitro using DNA substrates with different intervening distances (73-913 bp) between two directly oriented recombination (six) sites. All analyzed HMGB proteins (HMGB1 to HMGB5) could promote beta recombination, but depending on the DNA substrate with different efficiencies. The HMGB1 protein displayed an activity comparable to that of the natural promoting protein Hbsu, whereas the other HMGB proteins were less effective. Phosphorylation of the HMGB1 protein resulted in an increased efficiency of HMGB1 to promote beta recombination. Analyses of DNA substrates with closely spaced six sites demonstrated that in the presence of HMGB1 the recombination rate was correlated to the distance between the six sites, but independent of the helical orientation of the six sites. Using a Bacillus subtilis strain defective in Hbsu, the coexpression of beta recombinase and HMGB1 (or a truncated HMGB1 derivative) revealed that a plant HMG-box domain protein is sufficient for assisting beta to catalyze recombination in vivo. Our results using beta recombination as a model system suggest that the various plant HMGB proteins (and their posttranslationally modified versions) have the potential of forming a repertoire of different DNA structures, which is compatible with the idea that the HMGB proteins can act as architectural factors in a variety of nucleoprotein structures.
Stemmer
Christian
C
Department of Biotechnology, Institute of Life Sciences, Aalborg University, Sohngaardsholmsvej 49, DK-9000 Aalborg, Denmark.
Fernández
Silvia
S
Lopez
Gema
G
Alonso
Juan C
JC
Grasser
Klaus D
KD
D001412:Bacillus subtilis / Q000737:chemistry* / Q000235:genetics; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins / Q000172:deficiency / Q000235:genetics; D024742:HMG-Box Domains / Q000235:genetics; D024002:HMGB Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D024243:HMGB1 Protein / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry / Q000235:genetics; D011995:Recombination, Genetic; D003313:Zea mays / Q000737:chemistry / Q000235:genetics
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D024002:HMGB Proteins; D024243:HMGB1 Protein; C075779:Hbsu protein, bacteria; D010940:Plant Proteins