Translational efficiency of rpoS mRNA from Borrelia burgdorferi: Effects of the length and sequence of the mRNA leader region

被引：7

作者：

Archambault, Linda ^{[1
]}

Linscott, Joshua ^{[1
]}

Swerdlow, Nicholas ^{[1
]}

Boyland, Kathleen ^{[1
]}

Riley, Eammon ^{[1
]}

Schlax, Paula ^{[1
]}

机构：

[1] Bates Coll, Program Biol Chem, Lewiston, ME 04240 USA

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 2013年 / 433卷 / 01期

基金：

美国国家卫生研究院;

关键词：

Borrelia burgdorferi; Translational regulation; rpoS; COLI ALPHA-OPERON; LYME-DISEASE; EXPRESSION; TEMPERATURE; REPRESSION; PH; IDENTIFICATION; INITIATION; PROTEIN;

D O I：

10.1016/j.bbrc.2013.02.063

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Regulation of the enzootic cycle in Borrelia burgdorferi requires a shift to the RNA polymerase alternative sigma factor, RpoS. We used in vitro and in vivo assays to assess the relative importance of the putative Shine-Dalgarno sequence and its sequestration for the translational efficiency of rpoS. We created mutant leader regions in which we either removed the Shine-Dalgarno sequence, disrupted the secondary structure or both. Binding assays and toeprint assays demonstrated that both the presence and the availability of the Shine-Dalgarno sequence are important to the efficiency and specificity of ribosome binding. Adding a DsrA(Bb) mimic in the form of a single-stranded DNA oligonucleotide increased the level and specificity of binding ribosomes to the transcript with an extended leader, presumably by making the Shine-Dalgarno sequence available for binding. In in vivo assays we confirmed that the Shine-Dalgarno sequence must be both present and un-sequestered in order for translation to proceed efficiently. The longer transcript was significantly better translated in B. burgdorferi at 37 degrees C than at 26 degrees C, lending support to the hypothesis that DsrA(Bb) acts as a temperature-dependent stimulator of translation. These studies demonstrate that translational regulation of gene expression in B. burgdorferi may be an important mechanism for responding to environmental signals important in the enzootic cycle. (c) 2013 Elsevier Inc. All rights reserved.

引用

页码：73 / 78

页数：6

共 32 条

[21] Borrelia burgdorferi proteins whose expression is similarly affected by culture temperature and pH [J].

Ramamoorthy, R ;

Scholl-Meeker, D .

INFECTION AND IMMUNITY, 2001, 69 (04) :2739-2742

[22] Gene Regulation in Borrelia burgdorferi [J].

Samuels, D. Scott .

ANNUAL REVIEW OF MICROBIOLOGY, VOL 65, 2011, 65 :479-499

[23] Translational repression mechanisms in prokaryotes [J].

Schlax, PJ ;

Worhunsky, DJ .

MOLECULAR MICROBIOLOGY, 2003, 48 (05) :1157-1169

[24] Translational repression of the Escherichia coli α operon mRNA -: Importance of an mRNA conformational switch and a ternary entrapment complex [J].

Schlax, PJ ;

Xavier, KA ;

Gluick, TC ;

Draper, DE .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (42) :38494-38501

[25] INDUCTION OF AN OUTER SURFACE PROTEIN ON BORRELIA-BURGDORFERI DURING TICK FEEDING [J].

SCHWAN, TG ;

PIESMAN, J ;

GOLDE, WT ;

DOLAN, MC ;

ROSA, PA .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1995, 92 (07) :2909-2913

[26]

SHINE J, 1974, Proceedings of the National Academy of Sciences of the United States of America, V71, P1342, DOI 10.1073/pnas.71.4.1342

[27]

Singh SK, 2004, LANCET INFECT DIS, V4, P575

[28]

Skare J.T., 2010, GENE REGUL TRANSCRIP, P67

[29] ALLOSTERIC MECHANISM FOR TRANSLATIONAL REPRESSION IN THE ESCHERICHIA-COLI ALPHA-OPERON [J].

SPEDDING, G ;

DRAPER, DE .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (10) :4399-4403

[30] THE SPIROCHETAL ETIOLOGY OF LYME-DISEASE [J].

STEERE, AC ;

GRODZICKI, RL ;

KORNBLATT, AN ;

CRAFT, JE ;

BARBOUR, AG ;

BURGDORFER, W ;

SCHMID, GP ;

JOHNSON, E ;

MALAWISTA, SE .

NEW ENGLAND JOURNAL OF MEDICINE, 1983, 308 (13) :733-740

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