Isolation and characterization of the aspartokinase and aspartate semialdehyde dehydrogenase operon from mycobacteria

Jeffrey D. Cirillo, Torin Weisbrod, Lisa Pascopella, Barry R. Bloom, William R. Jacobs

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Diaminopimelic acid (DAP) is a major component of the peptidoglycan layer of the mycobacterial cell wall. The mycobacterial cell wall has been implicated as a potential virulence factor and is highly immunogenic. The pathway for biosynthesis of DAP may serve as a target in the design of antimycobacterial agents and construction of in vivo selection systems. Despite its significance, this biosynthetic pathway is poorly understood in mycobacteria. In order to develop a better understanding of mycobacterial DAP biosynthesis, the aspartate semialdehyde dehydrogenase (asd) genes of Mycobacterium smegmatis, bacille Calmete-Guerin (BCG), Mycobacterium avium, Mycobacterium leprae, and Mycobacterium tuberculosis were isolated. The M. smegmatis asd gene was isolated by complementation in Escherichia coli. This gene was then used to isolate the asd genes from other mycobacteria. The asd- complementing fragments from BCG and M. smegmatis were sequenced. An open reading frame upstream of the mycobacterial asd gene was identified as the mycobacterial aspartokinase gene (ask). Primer extension analysis revealed that the only transcriptional start in this region is found 5 of the ask gene. This observation indicates that the mycobacterial ask and asd genes are in an operon.

Original languageEnglish (US)
Pages (from-to)629-639
Number of pages11
JournalMolecular Microbiology
Volume11
Issue number4
DOIs
StatePublished - Feb 1994

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Aspartate-Semialdehyde Dehydrogenase
Aspartate Kinase
Operon
Mycobacterium
Diaminopimelic Acid
Mycobacterium smegmatis
Genes
Cell Wall
Mycobacterium avium
Mycobacterium leprae
Peptidoglycan
Biosynthetic Pathways
Virulence Factors
Mycobacterium tuberculosis
Open Reading Frames

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

Cite this

Isolation and characterization of the aspartokinase and aspartate semialdehyde dehydrogenase operon from mycobacteria. / Cirillo, Jeffrey D.; Weisbrod, Torin; Pascopella, Lisa; Bloom, Barry R.; Jacobs, William R.

In: Molecular Microbiology, Vol. 11, No. 4, 02.1994, p. 629-639.

Research output: Contribution to journalArticle

Cirillo, Jeffrey D. ; Weisbrod, Torin ; Pascopella, Lisa ; Bloom, Barry R. ; Jacobs, William R. / Isolation and characterization of the aspartokinase and aspartate semialdehyde dehydrogenase operon from mycobacteria. In: Molecular Microbiology. 1994 ; Vol. 11, No. 4. pp. 629-639.
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