A genetic and biochemical study of histidine biosynthesis in Micrococcus luteus

Abstract

Histidine auxotrophs of Micrococcus luteus strain ATCC 27141 were induced by treatment of the parent strain with N-methyl-N'-nitro-N-nitrosoguanidine. Auxotrophs were biochemically characterized by examining culture accumulations of histidine intermediates, using paper chromatography and the Bratton-Marshall test, and growth responses to L-histidinol. his(IG) mutants failed to accumulate Pauly-positive imidazoles; his(EAHF) mutants accumulated 5-amino-1-ribosyn-4-imidazole carboxamide; hisB mutants accumulated imidazoleglycerol; hisC mutants accumulated imidazoleacetol; hisD mutants, but did stimulate all other histidine mutants, blocked at earlier steps in the biosynthetic pathway. In addition, imidazoleglycerol phosphate dehydrase activity was assayed in representative mutants of each class. hisB mutants lacked activity for this enzyme.--Two -point, three-point, and cotransformation analyses resolved linkage relationships of histidine genes and in two gene clusters aided in determining their sequences. Histidine biosynthetic genes exist in at least four separate, unlinked regions of the chromosome. One histidine gene cluster linked to a tryptophan gene cluster and appears to be contiguous in the sequence his(IG) -his(EAHF)-trpE-trpC-trpA. A second and unlinked histidine cluster has the tentative gene sequence his(EAHF)-hisB-hisC-his(EAHF). The hisD gene and an unclassified mutant site his-94 are not linked to any of the other histidine genes examined in this study or to each other.