amiA is a negative regulator of acetamidase expression in Mycobacterium smegmatis

被引：24

作者：

Parish T. ^{[1
]}

Turner J. ^{[2
]}

Stoker N.G.

机构：

[1] Department of Medical Microbiology, Barts and the London, Qu. Mary's Sch. of Med. and Dent., London

[2] Dept. of Infectious and Trop. Dis., London Sch. of Hyg. and Trop. Med., London

来源：

BMC Microbiology | / 1卷 / 1期

关键词：

Acetamide; Inducible Promoter; Mycobacterium Smegmatis; Promoter Probe Vector; AmiE Expression;

D O I：

10.1186/1471-2180-1-19

中图分类号：

学科分类号：

摘要：

Background: The acetamidase of Mycobacterium smegmatis is a highly inducible enzyme. Expression of this enzyme is increased 100-fold when the substrate acetamide is present. The acetamidase gene is found immediately downstream of three open reading frames. Two of these are proposed to be involved in regulation. Results: We constructed a deletion mutant in one of the upstream ORFs (amiA). This mutant (Mad1) showed a constitutively high level of acetamidase expression. We identified four promoters in the upstream region using a β-galactosidase reporter gene. One of these (P2) was inducible in the wild-type, but was constitutively active in Mad1. Conclusions: These results demonstrate that amiA encodes a negative regulatory protein which interacts with P2. Since amiA has homology to DNA-binding proteins, it is likely that it exerts the regulatory effect by binding to the promoter to prevent transcription.

引用

页码：1 / 5

页数：4

共 13 条

[1]

Draper P., The aliphatic acylamide amidohydrolase of Mycobacterium smegmatis: Its inducible nature and relation to acyl-transfer to hydroxylamine, J. Gen. Microbiol., 46, pp. 111-123, (1967)

[2]

Mahenthiralingam E., Draper P., Davis E.O., Colston M.J., Cloning and sequencing of the gene which encodes the highly inducible acetamidase of Mycobacterium smegmatis, J. Gen. Microbiol., 139, pp. 575-583, (1993)

[3]

Parish T., Mahenthiralingam E., Draper P., Davis E.O., Colston M.J., Regulation of the inducible acetamidase gene of Mycobacterium smegmatis, Microbiol., 143, pp. 2267-2276, (1997)

[4]

Bashyam M.D., Kaushal D., Dasgupta S.K., Tyagi A.K., A study of the mycobacterial transcriptional apparatus: Identification of novel features in promoter elements, J. Bacteriol., 178, pp. 4847-4853, (1996)

[5]

Mulder M.A., Zappe H., Steyn L.M., Mycobacterial promoters, Tubercle and Lung Disease, 78, pp. 211-223, (1997)

[6]

Mills J., Wyborn N.R., Greenwood J.A., Williams S.G., Jones C.W., Characterisation of a binding-protein-dependent, active transport system for short-chain amides and urea in the methylotrophic bacterium Methylophilus methylotrophus, Eur. J. Biochem., 251, pp. 45-53, (1998)

[7]

Payton M., Auty R., Delgoda R., Everett M., Sim E., Cloning and characterization of arylamine N-acetyltransferase genes from Mycobacterium smegmatis and Mycobacterium tuberculosis: Increased expression results in isoniazid resistance, J. Bacteriol., 181, pp. 1343-1347, (1999)

[8]

Triccas J.A., Parish T., Britton W.J., Gicquel B., An inducible expression system permitting the efficient purification of a recombinant antigen from Mycobacterium smegmatis, FEMS Microbiol. Lett., 167, pp. 151-156, (1998)

[9]

Manabe Y.C., Chen J.M., Ko C.G., Chen P., Bishai W.R., Conditional sigma factor expression, using the inducible acetamidase promoter, reveals that the Mycobacterium tuberculosis sigF gene modulates expression of the 16-kilodalton alpha-crystallin homologue, J. Bacteriol., 181, pp. 7629-7633, (1999)

[10]

Parish T., Stoker N.G., Use of a flexible cassette method to generate a double unmarked Mycobacterium tuberculosis tlyA plcABC mutant by gene replacement, Microbiol., 146, pp. 1969-1975, (2000)

← 1 2 →