Distance and end configuration effects on VDJP-mediated DNA joining.
We have previously reported the cloning of a protein, VDJP, that is capable of binding the nonamer element of the V(D)J Recombinational Signal Sequence (RSS) as well as joining linear DNA fragments containing RSS elements in vitro. We show here that the linearized DNA molecules must contain a 5' extension or blunt end in order to be joined by VDJP. DNAs with 3' extensions are not efficiently joined by VDJP. Furthermore, the joining activity of DNAs with 5' extensions is significantly increased as the distance between the end and the RSS decreases. It is not yet clear what role VDJP plays in vivo, because our assay may not mimic exactly the in vivo DNA intermediates.
Guilliams
T G
TG
Department of Microbiology, Medical College of Wisconsin, Milwaukee 53226.
Teng
M
M
Halligan
B D
BD
D000818:Animals; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D004254:DNA Nucleotidyltransferases / Q000378:metabolism*; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase; D015252:Deoxyribonucleases, Type II Site-Specific; D007144:Immunoglobulin J-Chains / Q000235:genetics; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D046911:Macromolecular Substances; D051379:Mice; D010957:Plasmids; D021382:Protein Sorting Signals / Q000378:metabolism; D015183:Restriction Mapping; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D045567:VDJ Recombinases; D014764:Viral Proteins / Q000378:metabolism
D007144:Immunoglobulin J-Chains; D007145:Immunoglobulin kappa-Chains; D046911:Macromolecular Substances; D021382:Protein Sorting Signals; D014764:Viral Proteins; C069983:gene 43 protein, Enterobacteria phage T4; D004247:DNA; D004254:DNA Nucleotidyltransferases; D045567:VDJ Recombinases; D004259:DNA-Directed DNA Polymerase; D004262:DNA Restriction Enzymes; C024388:CTGCAG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific