A Conserved Tetranucleotide Repeat is Necessary for Wild-type Expression of the Moraxella Catarrhalis UspA2 Protein

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 2006 Sep 12

PMID 16963572

Citations 15

Authors

Ahmed S Attia

Eric J Hansen

Affiliations

Soon will be listed here.

Abstract

The UspA2 protein has been shown to be directly involved in the serum-resistant phenotype of Moraxella catarrhalis. The predicted 5'-untranslated regions (UTR) of the uspA2 genes in several different M. catarrhalis strains were shown to contain various numbers (i.e., 6 to 23) of a heteropolymeric tetranucleotide (AGAT) repeat. Deletion of the AGAT repeats from the uspA2 genes in the serum-resistant M. catarrhalis strains O35E and O12E resulted in a drastic reduction in UspA2 protein expression and serum resistance. PCR and transformation were used to construct a series of M. catarrhalis O12E strains that differed only in the number of AGAT repeats in their uspA2 genes. Expression of UspA2 was maximal in the presence of 18 AGAT repeats, although serum resistance attained wild-type levels in the presence of as few as nine AGAT repeats. Increased UspA2 expression was correlated with both increased binding of vitronectin and decreased binding of polymerized C9. Real-time reverse transcription-PCR analysis showed that changes in the number of AGAT repeats affected the levels of uspA2 mRNA, with 15 to 18 AGAT repeats yielding maximal levels. Primer extension analysis indicated that these AGAT repeats were contained in the 5'-UTR of the uspA2 gene. The mRNA transcribed from a uspA2 gene containing 18 AGAT repeats was found to have a longer half-life than that transcribed from a uspA2 gene lacking AGAT repeats. These data confirm that the presence of the AGAT repeats in the 5'-UTR of the uspA2 gene is necessary for both normal expression of the UspA2 protein and serum resistance.

Citing Articles

Moraxella catarrhalis phase-variable loci show differences in expression during conditions relevant to disease.

Tan A, Blakeway L, Taha , Yang Y, Zhou Y, Atack J PLoS One. 2020; 15(6):e0234306.

PMID: 32555615 PMC: 7302503. DOI: 10.1371/journal.pone.0234306.

The Moraxella catarrhalis phase-variable DNA methyltransferase ModM3 is an epigenetic regulator that affects bacterial survival in an in vivo model of otitis media.

Blakeway L, Tan A, Jurcisek J, Bakaletz L, Atack J, Peak I BMC Microbiol. 2019; 19(1):276.

PMID: 31818247 PMC: 6902483. DOI: 10.1186/s12866-019-1660-y.

Persistence of Moraxella catarrhalis in Chronic Obstructive Pulmonary Disease and Regulation of the Hag/MID Adhesin.

Murphy T, Brauer A, Pettigrew M, Lafontaine E, Tettelin H J Infect Dis. 2018; 219(9):1448-1455.

PMID: 30496439 PMC: 6467191. DOI: 10.1093/infdis/jiy680.

Potential impact of a Moraxella catarrhalis vaccine in COPD.

Perez A, Murphy T Vaccine. 2017; 37(37):5551-5558.

PMID: 28185742 PMC: 5545157. DOI: 10.1016/j.vaccine.2016.12.066.

Abundant Intergenic TAACTGA Direct Repeats and Putative Alternate RNA Polymerase β' Subunits in Marine Beggiatoaceae Genomes: Possible Regulatory Roles and Origins.

MacGregor B Front Microbiol. 2016; 6:1397.

PMID: 26733950 PMC: 4679880. DOI: 10.3389/fmicb.2015.01397.

References

Henderson I, Cappello R, Nataro J . Autotransporter proteins, evolution and redefining protein secretion. Trends Microbiol. 2000; 8(12):529-32. DOI: 10.1016/s0966-842x(00)01853-9. View

Murphy G, Connell T, Barritt D, Koomey M, Cannon J . Phase variation of gonococcal protein II: regulation of gene expression by slipped-strand mispairing of a repetitive DNA sequence. Cell. 1989; 56(4):539-47. DOI: 10.1016/0092-8674(89)90577-1. View

Bayliss C, Field D, Moxon E . The simple sequence contingency loci of Haemophilus influenzae and Neisseria meningitidis. J Clin Invest. 2001; 107(6):657-62. PMC: 208953. DOI: 10.1172/JCI12557. View

Faden H . The microbiologic and immunologic basis for recurrent otitis media in children. Eur J Pediatr. 2001; 160(7):407-13. DOI: 10.1007/s004310100754. View

Forsgren A, Brant M, Mollenkvist A, Muyombwe A, Janson H, Woin N . Isolation and characterization of a novel IgD-binding protein from Moraxella catarrhalis. J Immunol. 2001; 167(4):2112-20. DOI: 10.4049/jimmunol.167.4.2112. View

Verduin C, Hol C, Fleer A, van Dijk H, van Belkum A . Moraxella catarrhalis: from emerging to established pathogen. Clin Microbiol Rev. 2002; 15(1):125-44. PMC: 118065. DOI: 10.1128/CMR.15.1.125-144.2002. View

Seib K, Peak I, Jennings M . Phase variable restriction-modification systems in Moraxella catarrhalis. FEMS Immunol Med Microbiol. 2002; 32(2):159-65. DOI: 10.1111/j.1574-695X.2002.tb00548.x. View

Horton R, Hunt H, Ho S, Pullen J, Pease L . Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. Gene. 1989; 77(1):61-8. DOI: 10.1016/0378-1119(89)90359-4. View

Weiser J, Love J, Moxon E . The molecular mechanism of phase variation of H. influenzae lipopolysaccharide. Cell. 1989; 59(4):657-65. DOI: 10.1016/0092-8674(89)90011-1. View

10.

Weiser J, Maskell D, Butler P, Lindberg A, Moxon E . Characterization of repetitive sequences controlling phase variation of Haemophilus influenzae lipopolysaccharide. J Bacteriol. 1990; 172(6):3304-9. PMC: 209140. DOI: 10.1128/jb.172.6.3304-3309.1990. View

11.

CATLIN B . Branhamella catarrhalis: an organism gaining respect as a pathogen. Clin Microbiol Rev. 1990; 3(4):293-320. PMC: 358165. DOI: 10.1128/CMR.3.4.293. View

12.

Helminen M, Maciver I, Latimer J, Cope L, McCracken Jr G, Hansen E . A major outer membrane protein of Moraxella catarrhalis is a target for antibodies that enhance pulmonary clearance of the pathogen in an animal model. Infect Immun. 1993; 61(5):2003-10. PMC: 280795. DOI: 10.1128/iai.61.5.2003-2010.1993. View

13.

High N, Deadman M, Moxon E . The role of a repetitive DNA motif (5'-CAAT-3') in the variable expression of the Haemophilus influenzae lipopolysaccharide epitope alpha Gal(1-4)beta Gal. Mol Microbiol. 1993; 9(6):1275-82. DOI: 10.1111/j.1365-2958.1993.tb01257.x. View

14.

Fleischmann R, Adams M, White O, Clayton R, Kirkness E, Kerlavage A . Whole-genome random sequencing and assembly of Haemophilus influenzae Rd. Science. 1995; 269(5223):496-512. DOI: 10.1126/science.7542800. View

15.

Campagnari A, Ducey T, Rebmann C . Outer membrane protein B1, an iron-repressible protein conserved in the outer membrane of Moraxella (Branhamella) catarrhalis, binds human transferrin. Infect Immun. 1996; 64(9):3920-4. PMC: 174315. DOI: 10.1128/iai.64.9.3920-3924.1996. View

16.

Jennings M, Hood D, Peak I, Virji M, Moxon E . Molecular analysis of a locus for the biosynthesis and phase-variable expression of the lacto-N-neotetraose terminal lipopolysaccharide structure in Neisseria meningitidis. Mol Microbiol. 1995; 18(4):729-40. DOI: 10.1111/j.1365-2958.1995.mmi_18040729.x. View

17.

Aebi C, Maciver I, Latimer J, Cope L, STEVENS M, Thomas S . A protective epitope of Moraxella catarrhalis is encoded by two different genes. Infect Immun. 1997; 65(11):4367-77. PMC: 175628. DOI: 10.1128/iai.65.11.4367-4377.1997. View

18.

Aebi C, Cope L, Latimer J, Thomas S, Slaughter C, McCracken Jr G . Mapping of a protective epitope of the CopB outer membrane protein of Moraxella catarrhalis. Infect Immun. 1998; 66(2):540-8. PMC: 107939. DOI: 10.1128/IAI.66.2.540-548.1998. View

19.

Aebi C, Lafontaine E, Cope L, Latimer J, Lumbley S, McCracken Jr G . Phenotypic effect of isogenic uspA1 and uspA2 mutations on Moraxella catarrhalis 035E. Infect Immun. 1998; 66(7):3113-9. PMC: 108321. DOI: 10.1128/IAI.66.7.3113-3119.1998. View

20.

Chen D, Barniak V, VanDerMeid K, McMichael J . The levels and bactericidal capacity of antibodies directed against the UspA1 and UspA2 outer membrane proteins of Moraxella (Branhamella) catarrhalis in adults and children. Infect Immun. 1999; 67(3):1310-6. PMC: 96462. DOI: 10.1128/IAI.67.3.1310-1316.1999. View