THE RELATIONSHIP OF BACILLUS SUBTILIS PHYSIOLOGY AND HELICAL STRUCTURE AND ORGANIZATION (MACROFIBER, CELL SURFACE, HELIX HAND INVERSION).

Abstract

Helix hand inversion exhibited by Bacillus subtilis macrofibers is induced by changes in culture medium composition. The kinetics of this inversion are compared to those of temperature-induced inversions. D-alanine evokes a similar inversion process. The role of left-twist proteins(s), the existence of "memory", and the asymmetry of left to right versus right to left kinetics are confirmed within the context of these inversion regimes. Initiation time of right to left inversions is correlated to degree of pre-shift twist. Evidence is presented suggesting effective twist of the wall is defined by (1) the average of that twist conformation inserted prior to a shift in culture conditions and that of wall inserted following the shift and (2) the location of left-handed material within the wall. A constant 50 minute delay is observed before initiation of left to right inversions, irregardless of twist. Evidence is presented for a protein in the left to right inversion process. A classification system of macrofiber phenotypes based upon hand and degree of structural organization has been established. Three major classes are identified. Subclasses are shown to be distinguishable. Isotwist phenotypes of seven strains are defined upon a matrix of temperature and medium composition. These plots reveal a fundamental pattern of hand and organization that is present in each of the strains studied. The polarity of the four axes, the range of attainable twist conformations, and the existence of a right-hand maximum in the 12.5% SPl domain remain virtually constant. Major variations include extent of a disorganized band and/or the shifting of conformational range either left or right. Several mutants were transformed into A734, a strain that produces the tightest structures at all four matrix corners. Multiple mutations are responsible for the phenotypes of several strains. Evidence is presented for single genes that express as extreme left-handedness and stress at high temperature, swelling and stress in TB at high temperature, and reduction in structural organization produced in high TB content at low temperature.