geoscience.net logo
+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Interactions of the DNA polymerase X from African swine fever virus with gapped DNA substrates. Quantitative analysis of functional structures of the formed complexes


, : Interactions of the DNA polymerase X from African swine fever virus with gapped DNA substrates. Quantitative analysis of functional structures of the formed complexes. Biochemistry 46(45): 12909-12924

Energetics and specificity of interactions between the African swine fever virus polymerase X and gapped DNA substrates have been studied, using the quantitative fluorescence titration technique. Stoichiometries of pol X complexes, with the DNA substrates, are higher than suggested by NMR studies. This can be understood in the context of the functionally heterogeneous organization of the total DNA-binding site of pol X, which is composed of two DNA-binding subsites. The enzyme forms two different complexes with the gapped DNAs, differing dramatically in affinities. In the high-affinity complex, pol X engages the total DNA-binding site, forming the gap complex, while in the low-affinity the enzyme binds to the dsDNA parts of the gapped DNA, using only one of the DNA-binding subsites. As a result, the net number of ions released in the gap complex formation is significantly larger than in the binding of the dsDNA part. In the presence of Mg+2, pol X shows a strong preference for the ssDNA gaps having one and two nucleotides. Recognition of the short gaps already occurs in the ground state of the enzyme-DNA complex. Surprisingly, the specific structure necessary to recognize the short gaps is induced by magnesium binding to the enzyme. In the absence of Mg+2, pol X looses its selectivity for short ssDNA gaps. Pol X binds gapped DNAs with considerable cooperative interactions, which increase with the decreasing gap size. The functional implications of these findings for ASFV pol X activities are discussed.

US$19.90

PMID: 17941646

DOI: 10.1021/bi700677j


Other references

Meisinger, C.; Ryan, M.T.; Hill, K.; Model, K.; Lim, J.H.; Sickmann, A.; Müller, H.; Meyer, H.E.; Wagner, R.; Pfanner, N., 2001: Protein import channel of the outer mitochondrial membrane: a highly stable Tom40-Tom22 core structure differentially interacts with preproteins, small tom proteins, and import receptors. The preprotein translocase of the yeast mitochondrial outer membrane (TOM) consists of the initial import receptors Tom70 and Tom20 and a approximately 400-kDa (400 K) general import pore (GIP) complex that includes the central receptor Tom22, the...

Kapkaev A.R., 1985: Specific morphological changes in the placenta in chronic pyelonephritis. Meditsinskii Zhurnal Uzbekistana (10): 53-56

Wartenberg, L.T.ebusiewicz, B.K.rmanska, K., 1978: The effect of cyclophosphamide and streptomycin on the level of vitamin A in the livers of mice infected with Trichinella spiralis. Wiadomosci parazytologiczne4(6): 671-678

Ruggiero, C., 1975: Drip irrigation tests on tobacco. Irrigazione 22 (5) 3-7

Buck S.H.; Maurin Y.; Burks T.F.; Yamamura H.I., 1984: High affinity tritium labeled substance p binding to longitudinal muscle membranes of the guinea pig small intestine. The binding of 3H-substance P (3H-SP) to longitudinal muscle membranes of the guinea pig small intestine was characterized. The binding of 3H-SP exhibited a high affinity (Kd = 0.5 nM). It was saturable (Bmax = 2 fmol/mg tissue), reversible and te...

Suhonen, H.; Fernández, M.; Serimaa, R.; Suortti, P., 2005: Simulation of small-angle x-ray scattering from collagen fibrils and comparison with experimental patterns. Simulation of small-angle x-ray scattering from collagen in healthy and cancerous breast tissue may reveal detailed information on the structural changes in collagen. Collagen fibril is modelled as a cylinder with axially periodic step-function el...

Plaziat J.C., 1983: Ecologic and paleogeographic problems as basis of difficulty of stratigraphic correlations for beginning of Tertiary in Western Europe; discussion and propositions of correlation of Maestrichtian and Ilerdian. Newsletters on Stratigraphy 13(1): 1-20

Thresh, J.M., 1962: Abnormal black currant foliage caused by the gall mite Phytoptus ribis Nal. Conspicuous malformation of black currant leaves, sometimes referred to as "false" reversion, was associated with the presence of Eriophyid mites (Phytoptus ribis Nal.) in the apex of affected shoots. The symptom provides an indication o...

Liu, L.; Zhang, L-ming.; Zhag, X-cong., 2005: Junctional epithelialization of dental implants in animals. To detect the histological changes and the expression of proliferating cell nuclear antigen (PCNA) in junctional epithelialization of dental implants. The junctional epithelium was detected by HE staining and the PCNA expression was observed by im...

Nosé, Y., 2006: Food and Drug Administration workshop on regulatory process for pediatric mechanical circulatory support devices. Artificial Organs 30(6): 409-410