A chimeric actin carrying N-terminal portion of Tetrahymena actin does not bind to DNase I.
A chimeric actin gene was constructed from Tetrahymena actin sequence corresponding to residues 1-83 and Dictyostelium actin sequence corresponding to residues 84-375, and the gene was expressed in Dictyostelium cells. Using DNase I-affinity column, we revealed that the product of the chimeric actin gene was not retained in the column whereas intrinsic actin was retained. In conjunction with our previous data that Tetrahymena actin does not interact with DNase I [Hirono, M., Kumagai, Y., Numata, O., & Watanabe Y. (1989) Proc. Natl. Acad. Sci. U.S. 86, 75-79], we suggest that the binding site of DNase I in an ubiquitous actin is located in N-terminal region (residues 1-83).
Hirono
M
M
RIKEN Cell Bank, Institute of Physical and Chemical Research, Ibaraki, Japan.
Sutoh
K
K
Watanabe
Y
Y
Ohno
T
T
D000199:Actins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D002678:Chimera; D003850:Deoxyribonuclease I / Q000378:metabolism*; D004023:Dictyostelium / Q000235:genetics / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D005796:Genes; D011485:Protein Binding; D015183:Restriction Mapping; D013768:Tetrahymena / Q000235:genetics / Q000378:metabolism*; D014170:Transformation, Genetic
D000199:Actins; D003850:Deoxyribonuclease I