Bacteriology at UAB http://www.microbio.uab.edu/

• Streptococcus pneumoniae

• Mycobacterium tuberculosis

• Bacillus anthracis

• Mycoplasma

• biological mechanisms

• virulence factors

• pathogenesis

• vaccine development

• drug development

Bacteriology at UAB

Briles, David Streptococcus pneumoniae; vaccines

Hollingshead, Susan Microbial variation, streptococci

Nahm, Moon Vaccines; Streptococcus pneumoniae

Pritchard, David Glycobiology; streptococci

Wu, Hui Streptococci, glycoproteins, biofilms

Yother, Janet Capsules, streptococci

Streptococcus pneumoniae

Molecular mechanisms of virulence proteins of Streptococcus pneumoniae and their use as vaccine

antigens.

David E. Briles, PhDDepartments of Microbiology and Pediatrics

University of Alabama at Birmingham

PspA

coiled coil α-helix

proline-rich

choline-binding

PneumolysinAutolysinCell Membrane Cell Wall

Capsular Polysaccharide

Teichoic Acid

Neuraminidase A

IgA1 Protease

Lipoteichoic Acid

Phosphocholine

PspC / CbpA

Hyaluronidase

Protection-elicitingproteins are indicatedby colors

PneumococcalVirulence factors

PcpA

Financial support: NIH, Gates Foundation, PATH

Aims:

• to develop a pneumococcal vaccine based on a mixture

of pneumococcal protein antigens

• to develop a rapid test for pneumococcal infection

Janet Yother, Ph.D.Streptococcus pneumoniae capsular

polysaccharide synthesis and regulation

Cps2GCps2FCps2T

Cps2I

Membrane

UDP

Glc-1-P

Glc-6-P

GalU PGM

TDP

Rha

Glc

GlcUA

UDP

UDPTDP

Cps2L

Cps2K

Cps2NMO

Cps2J(Wzx)

Cps2H(Wzy)

Peptidoglycan

CytoplasmUnd-P

Cps2E

undecaprenyl phosphate

Released

BA C

D~P

dexB A B C D E T F G H I J K P L M N O aliA

Janet Yother, Ph.D.Streptococcus pneumoniae capsular polysaccharide

synthesis and regulation

INITIATION OLIGOSACCHARIDESYNTHESIS

DISSOCIATION“Low UDP-GlcUA”

EJECTIONFrequency / ChainLength Determinedby [UDP-GlcUA]

ENZYME?

High UDP-Glc

TRANSITION

HIGHLYPROCESSIVESYNTHESIS

“High” UDP-GlcUA

Synthase-dependent capsule synthesis

Moon NahmVaccine; S. pneumoniae; bacterial pathogenesis; immunity

Bratcher and Nahm. J Clin Microbiol. 2010.

Calix and Nahm. J Infect Dis. 2010. 202: 29-38.

Bratcher et al., Microbiology. 2010. 156: 555

Zartler et al. Carbohydr Res. 2009. 344:2586

Seo et al. Clin Vaccine Immunol. 2009. 16:1187.

Park et al. Infect Immun. 2009. 77:3374.

Bacteriology at UAB

Mycobacterium tuberculosis

Niederweis, Michael Outer membrane proteins of

M. tuberculosis

Steyn, Adrie Virulence of Mycobacterium tuberculosis

Transport across the outer membrane of M. tuberculosis

100 nm

100 nm

Uptake • sugars• lipids/fatty acids• ions (Fe2+, Cu2+)• drugs

Export

• lipids• sugars• Cu+

• drugs

Niederweis et al. (2010), Trends Microbiol. 18, 109-116

The Mycolab at UAB

Lab: 5 postdocs, 5 graduate students, 2 technicians

Financial support: $700,000 per year

• National Institutes of Health (NIH)

• Potts Memorial Foundation

• Center for AIDS Research of UAB

• Heiser Foundation, New York

Publications Faller et al. (2004). Science 303, 1189Hoffmann et al. (2008). Proc. Natl. Acad. Sci. USA 105, 3963Song et al. (2008). Tuberculosis 88, 526Danilchanka et al. (2008). Antimicrob. Agents Chemother. 52, 2503Butler et al. (2008). Proc. Natl. Acad. Sci. USA 105, 20647Huff et al. (2009). J. Biol. Chem 284, 10223

Science:

OMPs of Mtb: novel proteins, essential functions, medically important

Adrie SteynMechanism of Mycobacterium tuberculosis virulence

Singh et al. PLoS Pathog. 2009. e1000545

Kumar et al., J Biol Chem. 2008. 18032

Singh et al. Proc Natl Acad Sci U S A. 2007. 11562

Kumar et al. Proc Natl Acad Sci U S A. 2007. 11568

Bacteriology at UAB

Atkinson, Prescott Mycoplasma; Asthma

Dybvig, Kevin Mycoplasma; genetics

Higgins, N. Patrick Mobile DNA

Turnbough, Charles Bacillus anthracis, gene regulation

Other bacteria:

Kevin DybvigMycoplasmas; genetics; phenotypic switching;

DNA rearrangements

Simmons and Dybvig. FEMS Microbiol Lett. 2009. 295:77-81.

Daubenspeck et al. Mol Microbiol. 2009;72:1235-45

Luo et al. FEMS Microbiol Lett. 2009;290:195-8.

Luo et al. Infect Immun. 2008;76:4989-98.

Dybvig et al. Infect Immun. 2008;76:4000-8.

French et al. Mol Microbiol. 2008;69:67-76.

Structure and Function of the B. anthracis Exosporium—Turnbough Lab

• causes anthrax

• bioterroism agent

• infectious spores surrounded by an

exosporium

• exosporium required for spore viability

and infectivity

l ~20 exosporium proteins and glycoproteins are known. They are assembled into a complex structure via covalent linkages by novel mechanisms.

l Research focus: mechanisms and functions of exosporium protein covalent modifications (glycosylation, multi-site phosphorylation, isopeptide bond formation).

Bacillus anthraciscore

Recent Publications: Mol. Micro. 76:1527-1538 (2010) & J. Bacteriol. 192:1259-1268 (2010).

l With rare exceptions, in bacteria (e.g., E. coli and B. subtilis) the mechanisms that regulate expression of pyrimidine biosynthetic and salvage operons do not involve the participation of DNA-binding repressor or activator proteins.

l Instead, regulation occurs by mechanisms—or combinations of mechanisms— that rely entirely on the basic machinery of transcription or coupled transcription and translation. Such mechanisms include transcription attenuation, reiterative transcription, and/or transcription start-site switching.

l The Turnbough lab is working on the elucidation of these regulatory mechanisms at the molecular level and also on the description of new mechanisms of gene regulation without accessory proteins.

Regulation of Pyrimidine Gene Expression in Bacteria: Repression without Repressors—Turnbough Lab

Recent Reviews: Microbiol. Mol. Biol. Rev. 72:266-300 (2008) & Mol. Micro. 69:10-14 (2008).

l Example: Attenuation control of pyrG transcription via CTP-sensitve reiterative transcription in Bacillus subtilis. The pyrG gene encodes CTP synthetase, which produces CTP from UTP as the last step in pyrimidine nucleotide biosynthesis.