Purine biosynthesis is the bottleneck in trimethoprim‐treated Bacillus subtilis

Jennifer Janina Stepanek; Sina Schäkermann; Michaela Wenzel; Pascal Prochnow; Julia Elisabeth Bandow

doi:10.1002/prca.201600039

journal article Jul 21, 2016

Purine biosynthesis is the bottleneck in trimethoprim‐treated Bacillus subtilis

Jennifer Janina Stepanek Sina Schäkermann Michaela Wenzel

Pascal Prochnow Julia Elisabeth Bandow

PROTEOMICS – Clinical Applications Vol. 10 No. 9-10 pp. 1036-1048 · Wiley

View at Publisher Save 10.1002/prca.201600039

Abstract

PurposeTrimethoprim is a folate biosynthesis inhibitor. Tetrahydrofolates are essential for the transfer of C1 units in several biochemical pathways including purine, thymine, methionine, and glycine biosynthesis. This study addressed the effects of folate biosynthesis inhibition on bacterial physiology.Experimental designTwo complementary proteomic approaches were employed to analyze the response of Bacillus subtilis to trimethoprim. Acute changes in protein synthesis rates were monitored by radioactive pulse labeling of newly synthesized proteins and subsequent 2DE analysis. Changes in protein levels were detected using gel‐free quantitative MS.ResultsProteins involved in purine and histidine biosynthesis, the σB‐dependent general stress response, and sporulation were upregulated. Most prominently, the PurR‐regulon required for de novo purine biosynthesis was derepressed indicating purine depletion. The general stress response was activated energy dependently and in a subpopulation of treated cultures an early onset of sporulation was observed, most likely triggered by low guanosine triphosphate levels. Supplementation of adenosine triphosphate, adenosine, and guanosine to the medium substantially decreased antibacterial activity, showing that purine depletion becomes the bottleneck in trimethoprim‐treated B. subtilis.Conclusions and clinical relevanceThe frequently prescribed antibiotic trimethoprim causes purine depletion in B. subtilis, which can be complemented by supplementing purines to the medium.

Topics

No keywords indexed for this article. Browse by subject →

References

61

[1]

Review on Antimicrobial Resistance: Tackling a Crisis for the Health and Wealth of Nations.London 2014.

[2]

Gleckman R. "Drug therapy reviews: trimethoprim‐sulfamethoxazole" Am. J. Hosp. Pharm. (1979)

[3]

10.1002/bies.10114

[4]

10.1039/np9860300395

[5]

10.1002/9780470122723.ch9

[6]

10.1016/s0083-6729(08)60762-1

[7]

10.3390/nu3010118

[8]

10.1002/ange.19350484202

[9]

10.1136/jcp.21.2.202

[10]

10.1128/aac.04141-14

[11]

10.1128/aac.02282-13

[12]

10.1016/j.ijantimicag.2015.04.002

[13]

10.1128/aac.01658-08

[14]

10.1128/aac.00705-09

[15]

10.1051/vetres:2001123

[16]

10.1186/1471-2164-12-583

[17]

10.1128/aac.47.3.948-955.2003

[18]

10.1128/aac.00078-11

[19]

10.1128/aac.01380-12

[20]

10.1128/jb.149.1.391-394.1982

[21]

10.1128/jb.172.12.7211-7226.1990

[22]

10.1128/jb.81.5.741-746.1961

[23]

10.1099/00221287-139-9-2041

[24]

10.1002/1615-9861(200104)1:5<699::aid-prot699>3.0.co;2-c

[25]

10.1002/pmic.200701184

[26]

10.1002/pmic.200400907

[27]

Sambrook J. R. (2001)

[28]

10.1093/genetics/73.2.215

[29]

10.1128/jb.00107-13

[30]

10.1128/jb.185.15.4305-4314.2003

[31]

SigB-Dependent General Stress Response in Bacillus subtilis and Related Gram-Positive Bacteria

Michael Hecker, Jan Pané-Farré, Völker Uwe

Annual Review of Microbiology 10.1146/annurev.micro.61.080706.093445

[32]

10.1006/jmbi.1999.2665

[33]

10.1128/jb.183.21.6422-6428.2001

[34]

A PP2C phosphatase containing a PAS domain is required to convey signals of energy stress to the σB transcription factor of Bacillus subtilis

Kamni Vijay, Margaret S. Brody, Elisabeth Fredlund et al.

Molecular Microbiology 10.1046/j.1365-2958.2000.01697.x

[35]

10.1111/j.1574-6968.2001.tb10678.x

[36]

10.1128/jb.187.22.7554-7560.2005

[37]

10.1101/gad.874201

[38]

10.1128/jb.146.2.605-613.1981

[39]

10.1128/jb.151.2.1062-1065.1982

[40]

10.1016/0005-2787(77)90287-8

[41]

10.1073/pnas.57.1.114

[42]

10.1007/bf00338408

[43]

10.1128/jb.183.21.6175-6183.2001

[44]

10.1073/pnas.92.16.7455

[45]

Riboswitches Control Fundamental Biochemical Pathways in Bacillus subtilis and Other Bacteria

Maumita Mandal, Benjamin Boese, Jeffrey E Barrick et al.

Cell 10.1016/s0092-8674(03)00391-x

[46]

10.1128/jb.156.3.1107-1117.1983

[47]

10.1128/jb.05899-11

[48]

10.1128/jb.169.7.2977-2983.1987

[49]

10.1271/bbb.100063

[50]

10.1186/1471-2180-13-301

Showing 50 of 61 references

Cited By

24

Metabolic reconstruction reveals ATP salvage as a key response to trimethoprim treatment

Cailean Carter, Dipali Singh · 2026

iScience

Metrics

24

Citations

61

References

Details

Published: Jul 21, 2016
Vol/Issue: 10(9-10)
Pages: 1036-1048
License: View

Authors

J

Jennifer Janina Stepanek