Mutational Analysis Supports Three-Hairpin Model of Attenuator for Transcription Regulation of <i>ilvBN</i>C Operon in <i>Corynebacterium glutamicum</i>
The <i>ilvBNC</i> operon in <i>Corynebacterium glutamicum</i> encodes key enzymes for the biosynthesis of branched-chain amino acids (L-isoleucine, L-leucine, and L-valine). This operon has been studied for quite a long time, and it is assumed that three hairpin mRNA structur...
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2025-01-01
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| author | Ludmila E. Ryabchenko Igor I. Titov Tatyana E. Leonova Tatyana I. Kalinina Tatyana V. Gerasimova Marina E. Sheremetieva Nikolay A. Kolchanov Tamara M. Khlebodarova Alexander S. Yanenko |
| author_facet | Ludmila E. Ryabchenko Igor I. Titov Tatyana E. Leonova Tatyana I. Kalinina Tatyana V. Gerasimova Marina E. Sheremetieva Nikolay A. Kolchanov Tamara M. Khlebodarova Alexander S. Yanenko |
| author_sort | Ludmila E. Ryabchenko |
| collection | DOAJ |
| description | The <i>ilvBNC</i> operon in <i>Corynebacterium glutamicum</i> encodes key enzymes for the biosynthesis of branched-chain amino acids (L-isoleucine, L-leucine, and L-valine). This operon has been studied for quite a long time, and it is assumed that three hairpin mRNA structures can be formed in its regulatory region; however, their functionality and role in the attenuation mechanism of the <i>ilvBNC</i> operon are not completely clear. In the present work, we performed a mutational analysis of mRNA secondary structures in the regulatory region of the <i>C. glutamicum ilvBNC</i> operon, which allowed us to propose a model of the regulation of its transcription involving three mRNA hairpins that essentially act as a transcription terminator, an antiterminator, and an antiantiterminator. In this work, we proved the existence of a transcription terminator in this operon and experimentally confirmed the effectiveness of its influence on the expression of the <i>ilvBNC</i> operon, AHAS enzyme activity, and valine production. We demonstrated the unique functional features of this attenuator, which, due to the overlapping of the terminator and antiterminator hairpins, is capable of rapid low-energy transitions between them without the complete disruption of the hairpin structures. |
| format | Article |
| id | doaj-art-e5b5b19b658940e8adbaa2d1a1cc4c46 |
| institution | DOAJ |
| issn | 2076-2607 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj-art-e5b5b19b658940e8adbaa2d1a1cc4c462025-08-20T02:45:00ZengMDPI AGMicroorganisms2076-26072025-01-0113229110.3390/microorganisms13020291Mutational Analysis Supports Three-Hairpin Model of Attenuator for Transcription Regulation of <i>ilvBN</i>C Operon in <i>Corynebacterium glutamicum</i>Ludmila E. Ryabchenko0Igor I. Titov1Tatyana E. Leonova2Tatyana I. Kalinina3Tatyana V. Gerasimova4Marina E. Sheremetieva5Nikolay A. Kolchanov6Tamara M. Khlebodarova7Alexander S. Yanenko8National Research Center “Kurchatov Institute”, Kurchatov Genomic Center, Akademika Kurchatova pl. 1, 123182 Moscow, RussiaDepartment of Systems Biology, Institute of Cytology and Genetics SB RAS, Akademika Lavrentyev Ave., 10, 630090 Novosibirsk, RussiaNational Research Center “Kurchatov Institute”, Kurchatov Genomic Center, Akademika Kurchatova pl. 1, 123182 Moscow, RussiaNational Research Center “Kurchatov Institute”, Kurchatov Genomic Center, Akademika Kurchatova pl. 1, 123182 Moscow, RussiaNational Research Center “Kurchatov Institute”, Kurchatov Genomic Center, Akademika Kurchatova pl. 1, 123182 Moscow, RussiaNational Research Center “Kurchatov Institute”, Kurchatov Genomic Center, Akademika Kurchatova pl. 1, 123182 Moscow, RussiaDepartment of Systems Biology, Institute of Cytology and Genetics SB RAS, Akademika Lavrentyev Ave., 10, 630090 Novosibirsk, RussiaDepartment of Systems Biology, Institute of Cytology and Genetics SB RAS, Akademika Lavrentyev Ave., 10, 630090 Novosibirsk, RussiaNational Research Center “Kurchatov Institute”, Kurchatov Genomic Center, Akademika Kurchatova pl. 1, 123182 Moscow, RussiaThe <i>ilvBNC</i> operon in <i>Corynebacterium glutamicum</i> encodes key enzymes for the biosynthesis of branched-chain amino acids (L-isoleucine, L-leucine, and L-valine). This operon has been studied for quite a long time, and it is assumed that three hairpin mRNA structures can be formed in its regulatory region; however, their functionality and role in the attenuation mechanism of the <i>ilvBNC</i> operon are not completely clear. In the present work, we performed a mutational analysis of mRNA secondary structures in the regulatory region of the <i>C. glutamicum ilvBNC</i> operon, which allowed us to propose a model of the regulation of its transcription involving three mRNA hairpins that essentially act as a transcription terminator, an antiterminator, and an antiantiterminator. In this work, we proved the existence of a transcription terminator in this operon and experimentally confirmed the effectiveness of its influence on the expression of the <i>ilvBNC</i> operon, AHAS enzyme activity, and valine production. We demonstrated the unique functional features of this attenuator, which, due to the overlapping of the terminator and antiterminator hairpins, is capable of rapid low-energy transitions between them without the complete disruption of the hairpin structures.https://www.mdpi.com/2076-2607/13/2/291<i>Corynebacterium glutamicum</i>transcription regulationattenuation<i>ilvBNC</i> operonacetohydroxy acid synthase (AHAS) |
| spellingShingle | Ludmila E. Ryabchenko Igor I. Titov Tatyana E. Leonova Tatyana I. Kalinina Tatyana V. Gerasimova Marina E. Sheremetieva Nikolay A. Kolchanov Tamara M. Khlebodarova Alexander S. Yanenko Mutational Analysis Supports Three-Hairpin Model of Attenuator for Transcription Regulation of <i>ilvBN</i>C Operon in <i>Corynebacterium glutamicum</i> Microorganisms <i>Corynebacterium glutamicum</i> transcription regulation attenuation <i>ilvBNC</i> operon acetohydroxy acid synthase (AHAS) |
| title | Mutational Analysis Supports Three-Hairpin Model of Attenuator for Transcription Regulation of <i>ilvBN</i>C Operon in <i>Corynebacterium glutamicum</i> |
| title_full | Mutational Analysis Supports Three-Hairpin Model of Attenuator for Transcription Regulation of <i>ilvBN</i>C Operon in <i>Corynebacterium glutamicum</i> |
| title_fullStr | Mutational Analysis Supports Three-Hairpin Model of Attenuator for Transcription Regulation of <i>ilvBN</i>C Operon in <i>Corynebacterium glutamicum</i> |
| title_full_unstemmed | Mutational Analysis Supports Three-Hairpin Model of Attenuator for Transcription Regulation of <i>ilvBN</i>C Operon in <i>Corynebacterium glutamicum</i> |
| title_short | Mutational Analysis Supports Three-Hairpin Model of Attenuator for Transcription Regulation of <i>ilvBN</i>C Operon in <i>Corynebacterium glutamicum</i> |
| title_sort | mutational analysis supports three hairpin model of attenuator for transcription regulation of i ilvbn i c operon in i corynebacterium glutamicum i |
| topic | <i>Corynebacterium glutamicum</i> transcription regulation attenuation <i>ilvBNC</i> operon acetohydroxy acid synthase (AHAS) |
| url | https://www.mdpi.com/2076-2607/13/2/291 |
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