Escherichia coli mrdAts and mrdBts mutants forming spherical cells at 42C, were employed to investigate the possible role of both inner and outer membrance structures in the determination of cell shape of gram-negative cells. Spherical cells, but not rod-shaped wild types, were specifically killed by anionic detergents, such as sarkosyl, sodium dodecylsulfate and sodium deoxycholate. From the spherical intact cells grown overnight at 42C, much more proteins were released by sakosyl.