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J Bacteriol. 1971 March; 105(3): 1028-1035
Copyright © 1971 American Society for Microbiology. All Rights Reserved.

Mechanism of D-Cycloserine Action: Transport Mutants for D-Alanine, D-Cycloserine, and Glycine¹

^a Biochemistry Division, Department of Chemistry, Northwestern University, Evanston, Illinois 60201

ABSTRACT

The accumulation of D-alanine and the accumulation of glycine in Escherichia coli are related and appear to be separate from the transport of L-alanine. The analysis of four D-cycloserine-resistant mutants provides additional support for this conclusion. The first-step mutant from E. coli K-12 that is resistant to D-cycloserine was characterized by the loss of the high-affinity line segment of the D-alanine-glycine transport system in the Lineweaver-Burk plot. This mutation, which is linked to the met₁ locus, also resulted in the loss of the ability to transport D-cycloserine. The second-step mutation that is located 0.5 min from the first-step mutation resulted in the loss of the low-affinity line segment for the D-alanine-glycine transport system. The transport of L-alanine was decreased only 20 to 30% in each of these mutants. A multistep mutant from E. coli W that is 80-fold resistant to D-cycloserine lost >90% of the transport activity for D-alanine and glycine, whereas 75% of the transport activity for L-alanine was retained. E. coli W could utilize either D- or L-alanine as a carbon source, whereas the multistep mutant could only utilize L-alanine. Thus, a functioning transport system for D-alanine and glycine is required for both D-cycloserine action and growth on D-alanine.

² Present address: Kraftco Corp., Research and Development, Glenview, Ill.

³ National Science Foundation Summer Research Fellow, 1969.

⁴ Supported by Public Health Service Research Career Development Award 1-K3-AI-6950 from the National Institute of Allergy and Infectious Diseases.

¹ Presented in part at the 69th Annual Meeting of the American Society for Microbiology, Miami, Florida, 3–10 May, 1969. Taken from a thesis submitted by R. J. Wargel in partial fulfillment of the requirements for the Ph.D. degree from Northwestern University.

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