Involvement of the arginine repressor in lysine biosynthesis of Thermus thermophilus

Kei Fujiwara, Taishi Tsubouchi, Tomohisa Kuzuyama, Makoto Nishiyama

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Lysine biosynthesis of Thermus thermophilus proceeds in a similar way to arginine biosynthesis, and some lysine biosynthetic enzymes from T. thermophilus so far investigated have the potential to function in arginine biosynthesis. These observations suggest that arginine might regulate the expression of genes for lysine biosynthesis. To test this hypothesis, the argR gene encoding the regulator of arginine biosynthesis was cloned from T. thermophilus and its function in lysine biosynthesis was analysed. The addition of arginine to the culture medium inhibited the growth of an arginase gene knockout mutant of T. thermophilus, which presumably accumulates arginine inside the cells. Arginine-dependent growth inhibition was not alleviated by the addition of ornithine, which is a biosynthetic intermediate of arginine and serves as a peptidoglycan component of the cell wall in T. thermophilus. However, the growth inhibition was cancelled either by the simultaneous addition of lysine and ornithine or by a knockout of the argR gene, suggesting the involvement of argR in regulation of lysine biosynthesis in T thermophilus. Electrophoretic mobility shift assay and DNase I footprinting revealed that the ArgR protein specifically binds to the promoter region of the major lysine biosynthetic gene cluster. Furthermore, an α-galactosidase reporter assay for this promoter indicated that arginine repressed the promoter in an argR-dependent manner. These results indicate that lysine biosynthesis is regulated by arginine in T. thermophilus.

Original languageEnglish
Pages (from-to)3585-3594
Number of pages10
JournalMicrobiology
Volume152
Issue number12
DOIs
Publication statusPublished - 2006 Dec
Externally publishedYes

Fingerprint

Thermus thermophilus
Lysine
Arginine
Gene Knockout Techniques
Ornithine
Growth
Galactosidases
Arginase
Peptidoglycan
Deoxyribonuclease I
Electrophoretic Mobility Shift Assay
Regulator Genes
Multigene Family
Genetic Promoter Regions
Cell Wall
Culture Media

ASJC Scopus subject areas

  • Microbiology

Cite this

Involvement of the arginine repressor in lysine biosynthesis of Thermus thermophilus. / Fujiwara, Kei; Tsubouchi, Taishi; Kuzuyama, Tomohisa; Nishiyama, Makoto.

In: Microbiology, Vol. 152, No. 12, 12.2006, p. 3585-3594.

Research output: Contribution to journalArticle

Fujiwara, Kei ; Tsubouchi, Taishi ; Kuzuyama, Tomohisa ; Nishiyama, Makoto. / Involvement of the arginine repressor in lysine biosynthesis of Thermus thermophilus. In: Microbiology. 2006 ; Vol. 152, No. 12. pp. 3585-3594.
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