Hemantha Don Kulasekara

 

Education:

University of Colombo, Sri Lanka

Coppin State College, Baltimore, MD

Wake Forest University, Winston-Salem, NC

 

 

 

Post doctoral research: Stephen Lory Lab, Harvard Medical School: c-di-GMP signaling in Pseudomonas aeruginosa; regulation of Pseudomonas fimbriae by bvg like two-component regulators; HTS screening to discover inhibitors of SecA and Acp in Pseudomonas. Genomic evolution of the exoU island in Pseudomonas. 

Current Research:

1) Genes of unknown functions involved in membrane barrier function of Acinetobacter baumannii.

Using genetics and lipid biochemistry, we have identified number of previously uncharacterized genes that contribute to phospholipid metabolism in bacteria.

Past research:

1) Identified inhibitors of c-di-GMP mediated second messenger signaling by using both target-based and HTS-based drug discovery.

2) Characterized P. aeruginosa evolution in the CF airways (in collaboration with Dr. Lucas Hoffman lab).

3) Developed FRET-based biosensors to study second messenger dynamics in bacteria.

4) Studied the biological basis for inactivation of quorum sensing of Pseudomonas aeruginosa isolated from cystic fibrosis airways.

Research Interests:

Widefield and confocal microscopy, FRET-based biosensor technology, bacterial genetics, bacterial envelope, HTS screening/ antimicrobial discovery.

Publications:

1: Mayer-Hamblett N, Rosenfeld M, Gibson RL, Ramsey BW, Kulasekara HD,
Retsch-Bogart GZ, Morgan W, Wolter DJ, Pope CE, Houston LS, Kulasekara BR, Khan
U, Burns JL, Miller SI, Hoffman LR. Pseudomonas aeruginosa in vitro phenotypes
distinguish cystic fibrosis infection stages and outcomes. Am J Respir Crit Care
Med. 2014 Aug 1;190(3):289-97. doi: 10.1164/rccm.201404-0681OC. PubMed PMID:
24937177; PubMed Central PMCID: PMC4226041.

2: Mayer-Hamblett N, Ramsey BW, Kulasekara HD, Wolter DJ, Houston LS, Pope CE,
Kulasekara BR, Armbruster CR, Burns JL, Retsch-Bogart G, Rosenfeld M, Gibson RL,
Miller SI, Khan U, Hoffman LR. Pseudomonas aeruginosa phenotypes associated with
eradication failure in children with cystic fibrosis. Clin Infect Dis. 2014 Sep
1;59(5):624-31. doi: 10.1093/cid/ciu385. Epub 2014 May 26. PubMed PMID: 24863401;
PubMed Central PMCID: PMC4148602.

3: Kulasekara HD. Transposon mutagenesis. Methods Mol Biol. 2014;1149:501-19.
doi: 10.1007/978-1-4939-0473-0_39. PubMed PMID: 24818929.

4: Kulasekara BR, Kamischke C, Kulasekara HD, Christen M, Wiggins PA, Miller SI.
c-di-GMP heterogeneity is generated by the chemotaxis machinery to regulate
flagellar motility. Elife. 2013 Dec 17;2:e01402. doi: 10.7554/eLife.01402. PubMed
PMID: 24347546; PubMed Central PMCID: PMC3861689.

5: Jones C, Allsopp L, Horlick J, Kulasekara H, Filloux A. Subinhibitory
concentration of kanamycin induces the Pseudomonas aeruginosa type VI secretion
system. PLoS One. 2013 Nov 8;8(11):e81132. doi: 10.1371/journal.pone.0081132.
eCollection 2013. PubMed PMID: 24260549; PubMed Central PMCID: PMC3832665.

6: Pultz IS, Christen M, Kulasekara HD, Kennard A, Kulasekara B, Miller SI. The
response threshold of Salmonella PilZ domain proteins is determined by their
binding affinities for c-di-GMP. Mol Microbiol. 2012 Dec;86(6):1424-40. doi:
10.1111/mmi.12066. Epub 2012 Nov 19. PubMed PMID: 23163901.

7: Hocquet D, Llanes C, Thouverez M, Kulasekara HD, Bertrand X, Plésiat P, Mazel
D, Miller SI. Evidence for induction of integron-based antibiotic resistance by
the SOS response in a clinical setting. PLoS Pathog. 2012;8(6):e1002778. doi:
10.1371/journal.ppat.1002778. Epub 2012 Jun 14. PubMed PMID: 22719259; PubMed
Central PMCID: PMC3375312.

8: Mills E, Pultz IS, Kulasekara HD, Miller SI. The bacterial second messenger
c-di-GMP: mechanisms of signalling. Cell Microbiol. 2011 Aug;13(8):1122-9. doi:
10.1111/j.1462-5822.2011.01619.x. Epub 2011 Jun 24. Review. PubMed PMID:
21707905.

9: Christen M, Kulasekara HD, Christen B, Kulasekara BR, Hoffman LR, Miller SI.
Asymmetrical distribution of the second messenger c-di-GMP upon bacterial cell
division. Science. 2010 Jun 4;328(5983):1295-7. doi: 10.1126/science.1188658.
PubMed PMID: 20522779; PubMed Central PMCID: PMC3906730.

10: Hoffman LR, Richardson AR, Houston LS, Kulasekara HD, Martens-Habbena W,
Klausen M, Burns JL, Stahl DA, Hassett DJ, Fang FC, Miller SI. Nutrient
availability as a mechanism for selection of antibiotic tolerant Pseudomonas
aeruginosa within the CF airway. PLoS Pathog. 2010 Jan 8;6(1):e1000712. doi:
10.1371/journal.ppat.1000712. PubMed PMID: 20072604; PubMed Central PMCID:
PMC2795201.

11: Panchaud A, Scherl A, Shaffer SA, von Haller PD, Kulasekara HD, Miller SI,
Goodlett DR. Precursor acquisition independent from ion count: how to dive deeper
into the proteomics ocean. Anal Chem. 2009 Aug 1;81(15):6481-8. doi:
10.1021/ac900888s. PubMed PMID: 19572557; PubMed Central PMCID: PMC3086478.

12: Hoffman LR, Kulasekara HD, Emerson J, Houston LS, Burns JL, Ramsey BW, Miller
SI. Pseudomonas aeruginosa lasR mutants are associated with cystic fibrosis lung
disease progression. J Cyst Fibros. 2009 Jan;8(1):66-70. doi:
10.1016/j.jcf.2008.09.006. Epub 2008 Oct 29. PubMed PMID: 18974024; PubMed
Central PMCID: PMC2631641.

13: Scherl A, Shaffer SA, Taylor GK, Kulasekara HD, Miller SI, Goodlett DR.
Genome-specific gas-phase fractionation strategy for improved shotgun proteomic
profiling of proteotypic peptides. Anal Chem. 2008 Feb 15;80(4):1182-91. doi:
10.1021/ac701680f. Epub 2008 Jan 23. PubMed PMID: 18211032.

14: Holden N, Blomfield IC, Uhlin BE, Totsika M, Kulasekara DH, Gally DL.
Comparative analysis of FimB and FimE recombinase activity. Microbiology. 2007
Dec;153(Pt 12):4138-49. PubMed PMID: 18048927.

15: Kulasekara HD, Miller SI. Threonine phosphorylation times bacterial
secretion. Nat Cell Biol. 2007 Jul;9(7):734-6. PubMed PMID: 17603507.

16: D’Argenio DA, Wu M, Hoffman LR, Kulasekara HD, Déziel E, Smith EE, Nguyen H,
Ernst RK, Larson Freeman TJ, Spencer DH, Brittnacher M, Hayden HS, Selgrade S,
Klausen M, Goodlett DR, Burns JL, Ramsey BW, Miller SI. Growth phenotypes of
Pseudomonas aeruginosa lasR mutants adapted to the airways of cystic fibrosis
patients. Mol Microbiol. 2007 Apr;64(2):512-33. PubMed PMID: 17493132; PubMed
Central PMCID: PMC2742308.

17: Kulasekara BR, Kulasekara HD, Wolfgang MC, Stevens L, Frank DW, Lory S.
Acquisition and evolution of the exoU locus in Pseudomonas aeruginosa. J
Bacteriol. 2006 Jun;188(11):4037-50. PubMed PMID: 16707695; PubMed Central PMCID:
PMC1482899.

18: Kulasakara H, Lee V, Brencic A, Liberati N, Urbach J, Miyata S, Lee DG, Neely
AN, Hyodo M, Hayakawa Y, Ausubel FM, Lory S. Analysis of Pseudomonas aeruginosa
diguanylate cyclases and phosphodiesterases reveals a role for
bis-(3′-5′)-cyclic-GMP in virulence. Proc Natl Acad Sci U S A. 2006 Feb
21;103(8):2839-44. Epub 2006 Feb 13. PubMed PMID: 16477007; PubMed Central PMCID:
PMC1413825.

19: Kulasekara HD, Ventre I, Kulasekara BR, Lazdunski A, Filloux A, Lory S. A
novel two-component system controls the expression of Pseudomonas aeruginosa
fimbrial cup genes. Mol Microbiol. 2005 Jan;55(2):368-80. PubMed PMID: 15659157.

20: Sohanpal BK, Kulasekara HD, Bonnen A, Blomfield IC. Orientational control of
fimE expression in Escherichia coli. Mol Microbiol. 2001 Oct;42(2):483-94. PubMed
PMID: 11703669.

21: Kulasekara HD, Blomfield IC. The molecular basis for the specificity of fimE
in the phase variation of type 1 fimbriae of Escherichia coli K-12. Mol
Microbiol. 1999 Feb;31(4):1171-81. PubMed PMID: 10096084.

22: Blomfield IC, Kulasekara DH, Eisenstein BI. Integration host factor
stimulates both FimB- and FimE-mediated site-specific DNA inversion that controls
phase variation of type 1 fimbriae expression in Escherichia coli. Mol Microbiol.
1997 Feb;23(4):705-17. PubMed PMID: 9157242.