On the solution structure of the T4 sliding clamp (gp45).
Examination by time-resolved fluorescence spectroscopy of the trimeric bacteriophage T4 clamp protein labeled across its three subunit interfaces with a fluorescence resonance energy transfer (FRET) pair indicates that the clamp exists in just one state in solution, with one open and two closed interfaces. This is in contrast to what is observed in the X-ray crystal structure. The population distribution of the trFRET distance is bimodal, giving 67% as 17 A and 33% as 42 A. This leads to the conclusion that gp45 exists in an asymmetric open state in solution. The further increase in the separation of the FRET pair in the presence of the clamp loader and ATP may be ascribed to either further opening of the open interface or the opening of a closed interface. The ramifications for replisome remodeling by this pathway are discussed.
Department of Molecular Biology MB-19, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
David
D
Millar
Michael A
MA
Trakselis
Stephen J
SJ
Benkovic
D017122:Bacteriophage T4 / Q000737:chemistry* / Q000235:genetics; D018360:Crystallography, X-Ray; D031541:Fluorescence Resonance Energy Transfer; D008958:Models, Molecular; D017434:Protein Structure, Tertiary; D015534:Trans-Activators / Q000737:chemistry* / Q000235:genetics; D014364:Tryptophan / Q000737:chemistry; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics
D015534:Trans-Activators; D014764:Viral Proteins; C074555:gene 45 protein, Enterobacteria phage T4; D014364:Tryptophan