Skip to main page content (V体育官网)
U.S. flag

An official website of the United States government

Dot gov

The . gov means it’s official. Federal government websites often end in . gov or VSports app下载. mil. Before sharing sensitive information, make sure you’re on a federal government site. .

Https

The site is secure V体育官网. The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely. .

. 1999 Sep;67(9):4510-6.
doi: 10.1128/IAI.67.9.4510-4516.1999.

Stress-induced membrane association of the Streptococcus mutans GTP-binding protein, an essential G protein, and investigation of its physiological role by utilizing an antisense RNA strategy

Affiliations

Stress-induced membrane association of the Streptococcus mutans GTP-binding protein, an essential G protein, and investigation of its physiological role by utilizing an antisense RNA strategy

D Baev et al. Infect Immun. 1999 Sep.

"VSports注册入口" Abstract

SGP (for Streptococcus GTP-binding protein) is a Streptococcus mutans essential GTPase which has significant sequence identity to the previously identified Escherichia coli Era protein and to numerous other prokaryotic GTPase proteins of unknown function. Recent studies in our laboratory have addressed the possible role of SGP in the stress response of the oral pathogen S. mutans. Here we report that during growth in the early stationary phase, and in response to elevated temperatures or acidic pH, the distribution of SGP between the cytoplasm and the membranes of S. mutans cells varies. Immunoblot analysis of soluble and membrane protein fractions collected from the mid-log and early stationary growth phases of bacterial populations grown at normal temperature (37 degrees C) and at the elevated temperature of 43 degrees C, or at acidic pH, demonstrated that the total amount of SGP increased with the age of the bacterial culture, elevated temperature, or acidic pH. Furthermore, it was established that a substantial amount of SGP is associated with the membrane fraction under stress conditions. In order to investigate the physiological role of SGP, we constructed an S. mutans strain capable of chromosomal sgp antisense RNA expression, which interferes with the normal information processing of the sgp gene. Utilizing this strain, we determined conditions whereby the streptococcal cells can be depleted of SGP, thus avoiding the problem of constructing a conditional lethal system. From the results of measurements of the nucleotide pools extracted from the antisense strain and its isogenic counterpart, we propose that one of the physiological roles of SGP is regulation and modulation of the GTP/GDP ratio under different growth conditions. Moreover, we observed that in SGP-depleted cells the levels of glucan-binding protein A (GbpA) substantially increased, suggesting that GbpA may have stress response-related physiological functions VSports手机版. Finally, the potential applications of the antisense RNA approach that we employed are discussed. .

PubMed Disclaimer

Figures

FIG. 1
FIG. 1
Construction of S. mutans integration vector pSIV2. As described in the text, the final plasmid, pSIV2-SGPAN, was designed to express sgp antisense RNA under control of the scrB promoter.
FIG. 2
FIG. 2
(A) Immunoblot of membrane (M) and cytoplasmic (C) fractions from S. mutans SP2 grown in SMM with 1% glucose as the sole carbon source at 37°C to the mid-log growth phase (lanes 1 and 2) and to stationary phase (lanes 3 and 4). The bands corresponding to SGP and GAPDH are indicated with arrows. (B) Densitometric quantitation of the relative amounts of SGP in the membrane and cytoplasmic fractions.
FIG. 3
FIG. 3
Immunoblot of membrane (M) and cytoplasmic (C) fractions from S. mutans GS5(gtfB)::pSIV2 (lanes 1 and 2) and S. mutans GS5(gtfB)::pSIV2-SGPAN (lanes 3 and 4) grown in SMM–1% glucose at pH 5.50 and 37°C to the early stationary growth phase. The bands corresponding to SGP and GAPDH are indicated with arrows.
FIG. 4
FIG. 4
Immunoblot of membrane (M) and cytoplasmic (C) fractions from S. mutans GS5(gtfB)::pSIV2 (lanes 1 and 2) and S. mutans GS5(gtfB)::pSIV2-SGPAN (lanes 3 and 4) grown in SMM with 1% sucrose as the sole carbon source at 37°C to the early stationary growth phase. The bands corresponding to SGP and GAPDH are indicated with arrows.
FIG. 5
FIG. 5
Immunoblot of the total protein extracts from S. mutans GS5(gtfB)::pSIV2 (lane 1), S. mutans GS5(gtfB)::pSIV2-SGPAN (lane 2), and S. mutans BCH 150 (lane 3) grown in SMM–1% sucrose at 37°C to the early stationary growth phase. The bands corresponding to glucan-binding protein A (GbpA) and pyruvate kinase (PYK) are indicated with arrows.
FIG. 6
FIG. 6
Immunoblot of total protein extracts from S. mutans GS5(gtfB)::pSIV2-SGPAN grown in 5 ml of SMM with 1% sucrose (lane 1), 1% sucrose plus 0.1% glucose (lane 2), 1% sucrose plus 0.4% glucose (lane 3), and 1% glucose (lane 4) at 37°C to the early stationary growth phase. The bands corresponding to glucan-binding protein A (GbpA) and pyruvate kinase (PYK) are indicated with arrows.

V体育平台登录 - References

    1. Ahnn J, March P E, Takiff H E, Inouye M. A GTP-binding protein of Escherichia coli has homology to yeast RAS proteins. Proc Natl Acad Sci USA. 1986;83:8849–8853. - PMC - PubMed
    1. Banas J A, Potvin H C, Singh R N. The regulation of Streptococcus mutans glucan-binding protein A expression. FEMS Microbiol Lett. 1997;154:289–292. - PubMed
    1. Bourne H R, Sanders D A, McCormick F. The GTPase super-family: conserved structure and molecular mechanism. Nature (London) 1991;349:117–127. - PubMed
    1. Boyd D A, Cvitkovitch D G, Hamilton I R. Sequence, expression, and function of the gene for the nonphosphorylating, NADP-dependent glyceraldehyde-3-phosphate dehydrogenase of Streptococcus mutans. J Bacteriol. 1995;177:2622–2627. - VSports注册入口 - PMC - PubMed
    1. Cashel M, Gentry D R, Hernandez J, Vinella D. The stringent response. In: Neidhardt F C, Curtiss III R, Ingraham J L, Lin E C C, Low K B, Magasanik B, Reznikoff W S, Riley M, Schaechter M, Umbarger H E, editors. Escherichia coli and Salmonella: cellular and molecular biology. 2nd ed. Vol. 1. Washington, D.C: American Society for Microbiology; 1996. pp. 1458–1496.

Publication types

MeSH terms