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DNase1

A 33kd actin binding protein which also hydrolyses DNA. So far DNase 1, together with the ARP2/3 complex and tropomodulin  is amongst the few ABPs  to bind at the pointed end of actin filaments (Podolski & Steck, 1988). The physiological importance of the very tight association of DNAase 1 with actin is not certain as the two protein would not normally be expected to meet, however such interaction has been reported in some cell types at least (Malickablaszkiewicz, &  Roth,1983).  DNAase 1 has proved to be a valuable tool in the field as methods have been developed which utilize the fact that when DNAase 1 is bound by actin it no longer has the capacity to cleave DNA, in order to measure the G-actin content of actin solutions (Fox et al, 1981).  DNAase 1 played a vital part in the solution to the crystallographic structure of the actin monomer by preventing polymerization at high protein concentrations (Kabsch et al, 1990), the very high affinity of DNase 1 for actin was important in this regard.  DNase 1 has also been a very useful tool in the isolation of actin from various sources by affiinity chomatography (Schafer et al, 1998), although it does not bind to actin from Entamoeba (Gadasi, 1982).

The structure of DNase 1 (green) in complex with actin (blue) (Kabsch et al, 1990). 
Figure 1

DNase-1 actin binding

DNAse-1 binds G-actin with very high affinity () at the "top" of sub-domain 2 (Figure 1) and to the pointed end of microfilaments with a Kd of 1.9x10-9 and an association rate constant of ~1x106 (Podolski & Steck, 1988).  Not many other actin binding proteins bind to actin through this region (tropomodulin and the ARP2/3 complex) and competition between DNase-1 and other ABPs does not necessarily indicate competition for binding at this site since binding by DNase-1 affects even the C terminus of the actin monomer (Usmanova et al, 1997).  For example competition has been demonstrated between elongation factor 2 and DNase-1 but it remains to be seen why this is the case (Bektas et al, 1998).  DNase-1 also binds to some (Varma et al, 1987) but not all () actin related proteins.

DNase-1 and Apoptosis

Members of the DNase 1 family have been found to be be involved in DNA fragmentation during apoptosis

Protein Actin binding Reference
DNase I Yes
DNase II
DNase X
DNase g
DNAS1L2

References:-

Bektas, M., Nurten, R., Sayers, Z. & Bermek, E. (1998). “Interactions of elongation factor 2 with the cytoskeleton and interference with DNase I binding to actin.” Eur.J.Biochem. 256, 142-147.

Fox, J., Dockter, M.E. & Phillips, D. R. (1981). “An improved method for determining the actin filament content of non-muscle cells by the DNase-1 inhibition assay.” Analytical Biochemistry 117, 170-177.

Gadasi, H. (1982). “Isolated Entamoeba histolytica actin does not inhibit DNase-I activity.” Biochem. Biophys. Res. Comm. 104, 158-164.

Kabsch, W., Mannherz, H. G. Suck, D., Pai, E.F. & Holmes, K.C. (1990). “Atomic structure of the actin-DNase I complex.” Nature 347, 37-44.

Malickablaszkiewicz, M. &  Roth, J. S.  (1983). “Evidence for the presence of DNase actin complex in l1210 leukemia-cells.” FEBS Letters 153,  235-239.

Rao, K. (1981). "Actin polymerization, deoxyribonuclease and capping - a model for induction of DNA-synthesis in lymphocytes by concanavalin-a." Federation Proceedings 40: 1082.

Schafer, D. A.,  Jennings, P. B., & Cooper, J.A. (1998). “Rapid and efficient purification of actin from non-muscle sources.” Cell Mot.Cytoskel. 39, 166-171.

Shiokawa, D. & Tanuma, S.-i. (2001). "Characterization of human DNase I family endonucleases and activation of DNase g during apoptosis. Biochemistry 40, 143-152.

Ulmer, J. S., Herzka, A., Toy, K. J., Baker, D. L., Dodge, A. H., Sinicropi, D., Shak, S. & Lazarus, R. A. (1996) "Engineering actin-resistant human DNase1 for treatment of cystic fibrosis" PNAS. 93, 8225-8229.

Usmanova, A., Astier, C., Lebart, M.-C., Kwiatek, O., Papa, I., Boyer, M., Roustan, C. & Benyamin, Y. (1997) "Analysis of long-range structural effects induced by DNAse-I interaction with actin monomeric form or complexed to CapZ"., Biochimie. 79, 485-492.

Varma, M., Aebi, U., Fleming, J. & Leavitt, J. (1987) "A 60-kDa polypeptide in mammalian cells with epitopes related to actin" Expt.Cell Res. 173, 163-173

Weber, A.,  Pennise, C.R. & Pring, M. (1994). “DNase-I increases the rate-constant of depolymerization at the pointed (-) end of actin filament.” Biochemistry 33, 4780-4786.

Dalledonne, I., Milzani, A., Fascio, U., Ratti, A. & Colombo, R. (1993) Lithium preserves f-actin from the disarrangement induced by either dnase-i or cytochalasin-d, Biochemistry And Cell Biology Biochimie Et Biologie Cellulaire. 71, 440-446.

Grinstein, S. & Cohen, S. (1983) Measurement of sidedness of isolated plasma-membrane vesicles - quantitation of actin exposure by dnase-i inactivation, Analytical Biochemistry. 130, 151-157.

Heacock, C. S. & Bamburg, J. R. (1981) Application of the dnase i inhibition assay for quantitation of g-actin and f-actin in cultured-cells, Journal Of Cell Biology. 91.

Horono, M., Sutoh, K., Watanabe, Y. & Ohno, T. (1992) A chimaeric actin carrying N-terminal portion of Tetrahymena actin does not bind to DNase I, Biochem. Biophys. Res. Comm. 184, 1511-1516.

Kayalar, C., Örd, T., Testa, M. P., Zhong, L.-t. & Bredesen, D. E. (1996) Cleavage of actin by interleukin1b-converting enzyme to reverse DNase I inhibition, Proc. Natl. Acad. Sci. 93, 2234-2238.

Kiessling, P., Jahn, W., Maier, G., Polzar, B. & Mannherz, H. G. (1995) Purification and characterization of subtilisin cleaved actin lacking the segment of residues 43-47 in the DNase I binding loop, Biochemistry. 34, 14834-14842.

Malickablaszkiewicz, M. & Roth, J. S. (1981) Some factors affecting the interaction between actin in leukemic l1210 cells and dnase .1, Biochemical And Biophysical Research Communications. 102, 594-601.

Mannherz, H. G., Dieckhoff, J., Suck, D., Kabsch, W., Friebel, K. & Frimmer, M. (1985) Crystallization of cytoplasmic actin in complex with DNase-I and the interaction of dnase-i with polymeric actin, Journal Of Muscle Research And Cell Motility. 6, 77-78.

Podolski, J. L. & Steck, T. L. (1988) Association of deoxyribonuclease I with the pointed ends of actin filaments in human red blood cell membrane skeletons, J. Biol. Chem. 263, 638-645.

Podolski, J. L. & Steck, T. L. (1987) Deoxyribonuclease I association with erythrocyte-membrane actin-filaments, Federation Proceedings. 46, 1323.

Reitz, M., Knitza, R., Lanz, E. & Zahn, R. K. (1982) The effects of halothane on the dnase-i activity in an isolated enzyme preparation and in the dnase i-g actin complex, Chemico Biological Interactions. 42, 291-300.

Runge, M. R., Cooper, J. A., Tseng, P. & Pollard, T. D. (1982) Evaluation of acanthamoeba profilin binding to actin and actin-dnase by non-equilibrium dialysis, Biophysical Journal. 37.

Snabes, M. C., Boyd, A. E., Pardue, R. L. & Bryan, J. (1981) A DNase I-binding-immunoprecipitation assay for actin, J. Biol. Chem. 256, 6291-6295.

Weber, A., Pennise, C. R. & Pring, M. (1994) DNase-I increases the rate-constant of depolymerization at the pointed (-) end of actin filament, Biochemistry. 33, 4780-4786.

Zechel, K. (1980) Dissociation of the DNase I-actin complex by formamide, Eur. J. Biochem. 110, 337-341.

Zimmer, D. B., Pardue, R. L. & Goldstein, M. A. (1983) DNase I-actin interactions in the z-lattice of skeletal-muscle, Biophysical Journal. 41.

Kabsch, W., Mannherz, H. G., Suck, D., Pai, E. F. & Holmes, K. C. (1990) Atomic structure of the actin-DNase I complex, Nature. 347, 37-44.

Drummond, D. R., Hennessey, E. S. & Sparrow, J. C. (1992) The binding of mutant actins to profilin, ATP and DNase I, Eur.J.Biochem. 209, 171-179.

Jancso, A., Szilagyi, L., Wong, A. G. & Lu, R. C. (1984) Changes of lysine reactivities of actin in complex with DNase-1, Acta Biochim. Biophys. 19, 22.

Khaitlina, S., Moraczewska, J. & Strzelecka-Golaszewska, H. (1993) The actin/actin interactions involving the N-terminus of the DNase-I-binding loop the crucial for stabilization of the actin filament, Eur.J. Biochem. 218, 911-920.

Combeau, C. & Carlier, M.-F. (1992) Covalent modification of G-actin by pyridoxal 5'-phosphate: polymerization properties and interaction with DNase I and myosin subfragment 1, Biochemistry. 31, 300-309.

Enari, M., Sakahira, H., Yokoyama, H., Okawa, K., Iwamatsu, A. & Nagata, S. (1998) A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD, Nature. 391, 43-50.

Gadasi, H. (1982) Isolated Entamoeba-histolytica actin does not inhibit DNase-I activity, Biochem. Biophys. Res. Comm. 104, 158-164.

Blanchoin, L., Fievez, S., Travers, F., Carlier, M.-F. & Pantaloni, D. (1995) Kinetics of the interaction of myosin subfragment-1 with G-actin: effect of nucleotides and DNase I, J. Biol. chem. 270, 7125-7133.

 Ballweber, E., Giehl, K., Hannappel, E., Huff, T., Jockusch, B. M. & Mannhertz, H. G. (1998) Plant profilin induces actin polymerization from actin:b-thymosin complexes and competes directly with b-thymosins and with negative co-operativity with DNase I for binding to actin., FEBS letters. 425, 251-255.

Muncy, L. F. & Wolfe, J. S. (1981) Evidence for a non-actin inhibitor of deoxyribonuclease 1 (DNase-1) in Tetrahymena thermophila., J.Cell Biol. 91.

Leicher, A., Reitz, M., Knitza, R. & Lanz, E. (1982) Influence of halothane and enflurane on the activity of DNase1 and of the DNase-1 inhibitor G-actin in a cell-free system., Anaesthesist. 31, 513.

Los, M., Neubuser, D., Coy, J. F., Mozoluk, M., Poustka, A. & Schultze-Osthoff, K. (2000) Functional characterization of DNase X, a novel endonuclease expressed in muscle cells., Biochemistry. 39, 7365-7373.

Shiokawa, D. & Tanuma, S.-i. (2001) Characterization of human DNase 1 family endonucleases and activation of DNase g during apoptosis., Biochemistry. 40, 143-152.

Dobrowolski, J. M., Niesman, I. R. & Sibley, L. D. (1997) Actin in the parasite Toxoplasma gondii is encoded by a single copy gene, ACT1 and exists primarily in a globular form, Cell Mot.Cytoskel. 37, 253-262.

 Boyer, M., Roustan, C. & Benyamin, Y. (1985) DNaseI:actin complex - an immunological study, Bioscience Reports. 5, 39-46.

 
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