MECHANISM OF POST-SEGREGATIONAL KILLING - SECONDARY STRUCTURE-ANALYSIS OF THE ENTIRE HOK MESSENGER-RNA FROM PLASMID R1 SUGGESTS A FOLDBACK STRUCTURE THAT PREVENTS TRANSLATION AND ANTISENSE RNA-BINDING

T THISTED; NS SORENSEN; Kenn Gerdes

doi:10.1006/jmbi.1995.0186

MECHANISM OF POST-SEGREGATIONAL KILLING - SECONDARY STRUCTURE-ANALYSIS OF THE ENTIRE HOK MESSENGER-RNA FROM PLASMID R1 SUGGESTS A FOLDBACK STRUCTURE THAT PREVENTS TRANSLATION AND ANTISENSE RNA-BINDING

T THISTED, NS SORENSEN, Kenn Gerdes

34 Citationer (Scopus)

Abstract

The hok/sok system of plasmid R1 mediates plasmid stabilization by killing of plasmid-free cells. The Hok mRNA is very stable and can be translated into Hok killer protein. Translation of the Hok mRNA is inhibited by the small unstable Sok antisense RNA. Translation of hok is coupled to an overlapping reading frame termed mok. Translation of mok is tightly regulated by Sok RNA, and Sok RNA thus regulates hok translation indirectly through mok. The rapid decay of Sok RNA explains the onset of Hok synthesis in newborn plasmid-free segregants. However, a second control level is superimposed on this simple induction scheme, since the full-length Hok mRNA was found to be translationally inactive whereas a 3'-end truncated version of it was active. We have therefore previously suggested, that the 3'-terminal region of the full-length Hok mRNA encodes an element which prevents its translation. This element was termed fbi (fold-back inhibition). Here we describe the in vitro secondary structure of the entire Hok mRNA. Our results suggest a closed structure in which the 3'-end of the full-length Hok mRNA folds back onto the translational initiation region of mok. This structure explains why full-length Hok mRNA is translationally silent. The proposed structure was further supported by results obtained using mutations in the 3'-end fbi element. These `'structure closing'' mutations affected the structure much further upstream in the mok translational initiation region and concomitantly prevented antisense RNA binding to the same region of the mRNA. These results lend further support to the induction model that explains onset of Hok mRNA translation in plasmid-free segregants. The most important regulatory element in this model is the FBI structure formed between the 3'-end and the mok translational initiation region. This structure renders Hok mRNA translationally inactive and prevents antisense RNA binding, thus allowing the accumulation of a pool of mRNA which, by slow 3'-end processing, is activated in plasmid-free segregants, eventually leading to the elimination of these cells.

Originalsprog	Engelsk
Tidsskrift	Journal of Molecular Biology
Vol/bind	247
Udgave nummer	5
Sider (fra-til)	859-873
Antal sider	15
ISSN	0022-2836
DOI	https://doi.org/10.1006/jmbi.1995.0186
Status	Udgivet - 1995
Udgivet eksternt	Ja

Adgang til dokumentet

10.1006/jmbi.1995.0186

Citationsformater

MECHANISM OF POST-SEGREGATIONAL KILLING - SECONDARY STRUCTURE-ANALYSIS OF THE ENTIRE HOK MESSENGER-RNA FROM PLASMID R1 SUGGESTS A FOLDBACK STRUCTURE THAT PREVENTS TRANSLATION AND ANTISENSE RNA-BINDING. / THISTED, T; SORENSEN, NS; Gerdes, Kenn.

I: Journal of Molecular Biology, Bind 247, Nr. 5, 1995, s. 859-873.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

@article{a330e64435e44c04aae5c1300dac44b3,

title = "MECHANISM OF POST-SEGREGATIONAL KILLING - SECONDARY STRUCTURE-ANALYSIS OF THE ENTIRE HOK MESSENGER-RNA FROM PLASMID R1 SUGGESTS A FOLDBACK STRUCTURE THAT PREVENTS TRANSLATION AND ANTISENSE RNA-BINDING",

abstract = "The hok/sok system of plasmid R1 mediates plasmid stabilization by killing of plasmid-free cells. The Hok mRNA is very stable and can be translated into Hok killer protein. Translation of the Hok mRNA is inhibited by the small unstable Sok antisense RNA. Translation of hok is coupled to an overlapping reading frame termed mok. Translation of mok is tightly regulated by Sok RNA, and Sok RNA thus regulates hok translation indirectly through mok. The rapid decay of Sok RNA explains the onset of Hok synthesis in newborn plasmid-free segregants. However, a second control level is superimposed on this simple induction scheme, since the full-length Hok mRNA was found to be translationally inactive whereas a 3'-end truncated version of it was active. We have therefore previously suggested, that the 3'-terminal region of the full-length Hok mRNA encodes an element which prevents its translation. This element was termed fbi (fold-back inhibition). Here we describe the in vitro secondary structure of the entire Hok mRNA. Our results suggest a closed structure in which the 3'-end of the full-length Hok mRNA folds back onto the translational initiation region of mok. This structure explains why full-length Hok mRNA is translationally silent. The proposed structure was further supported by results obtained using mutations in the 3'-end fbi element. These `'structure closing'' mutations affected the structure much further upstream in the mok translational initiation region and concomitantly prevented antisense RNA binding to the same region of the mRNA. These results lend further support to the induction model that explains onset of Hok mRNA translation in plasmid-free segregants. The most important regulatory element in this model is the FBI structure formed between the 3'-end and the mok translational initiation region. This structure renders Hok mRNA translationally inactive and prevents antisense RNA binding, thus allowing the accumulation of a pool of mRNA which, by slow 3'-end processing, is activated in plasmid-free segregants, eventually leading to the elimination of these cells.",

author = "T THISTED and NS SORENSEN and Kenn Gerdes",

year = "1995",

doi = "10.1006/jmbi.1995.0186",

language = "English",

volume = "247",

pages = "859--873",

journal = "Journal of Molecular Biology",

issn = "0022-2836",

publisher = "Academic Press",

number = "5",

}

TY - JOUR

T1 - MECHANISM OF POST-SEGREGATIONAL KILLING - SECONDARY STRUCTURE-ANALYSIS OF THE ENTIRE HOK MESSENGER-RNA FROM PLASMID R1 SUGGESTS A FOLDBACK STRUCTURE THAT PREVENTS TRANSLATION AND ANTISENSE RNA-BINDING

AU - THISTED, T

AU - SORENSEN, NS

AU - Gerdes, Kenn

PY - 1995

Y1 - 1995

N2 - The hok/sok system of plasmid R1 mediates plasmid stabilization by killing of plasmid-free cells. The Hok mRNA is very stable and can be translated into Hok killer protein. Translation of the Hok mRNA is inhibited by the small unstable Sok antisense RNA. Translation of hok is coupled to an overlapping reading frame termed mok. Translation of mok is tightly regulated by Sok RNA, and Sok RNA thus regulates hok translation indirectly through mok. The rapid decay of Sok RNA explains the onset of Hok synthesis in newborn plasmid-free segregants. However, a second control level is superimposed on this simple induction scheme, since the full-length Hok mRNA was found to be translationally inactive whereas a 3'-end truncated version of it was active. We have therefore previously suggested, that the 3'-terminal region of the full-length Hok mRNA encodes an element which prevents its translation. This element was termed fbi (fold-back inhibition). Here we describe the in vitro secondary structure of the entire Hok mRNA. Our results suggest a closed structure in which the 3'-end of the full-length Hok mRNA folds back onto the translational initiation region of mok. This structure explains why full-length Hok mRNA is translationally silent. The proposed structure was further supported by results obtained using mutations in the 3'-end fbi element. These `'structure closing'' mutations affected the structure much further upstream in the mok translational initiation region and concomitantly prevented antisense RNA binding to the same region of the mRNA. These results lend further support to the induction model that explains onset of Hok mRNA translation in plasmid-free segregants. The most important regulatory element in this model is the FBI structure formed between the 3'-end and the mok translational initiation region. This structure renders Hok mRNA translationally inactive and prevents antisense RNA binding, thus allowing the accumulation of a pool of mRNA which, by slow 3'-end processing, is activated in plasmid-free segregants, eventually leading to the elimination of these cells.

AB - The hok/sok system of plasmid R1 mediates plasmid stabilization by killing of plasmid-free cells. The Hok mRNA is very stable and can be translated into Hok killer protein. Translation of the Hok mRNA is inhibited by the small unstable Sok antisense RNA. Translation of hok is coupled to an overlapping reading frame termed mok. Translation of mok is tightly regulated by Sok RNA, and Sok RNA thus regulates hok translation indirectly through mok. The rapid decay of Sok RNA explains the onset of Hok synthesis in newborn plasmid-free segregants. However, a second control level is superimposed on this simple induction scheme, since the full-length Hok mRNA was found to be translationally inactive whereas a 3'-end truncated version of it was active. We have therefore previously suggested, that the 3'-terminal region of the full-length Hok mRNA encodes an element which prevents its translation. This element was termed fbi (fold-back inhibition). Here we describe the in vitro secondary structure of the entire Hok mRNA. Our results suggest a closed structure in which the 3'-end of the full-length Hok mRNA folds back onto the translational initiation region of mok. This structure explains why full-length Hok mRNA is translationally silent. The proposed structure was further supported by results obtained using mutations in the 3'-end fbi element. These `'structure closing'' mutations affected the structure much further upstream in the mok translational initiation region and concomitantly prevented antisense RNA binding to the same region of the mRNA. These results lend further support to the induction model that explains onset of Hok mRNA translation in plasmid-free segregants. The most important regulatory element in this model is the FBI structure formed between the 3'-end and the mok translational initiation region. This structure renders Hok mRNA translationally inactive and prevents antisense RNA binding, thus allowing the accumulation of a pool of mRNA which, by slow 3'-end processing, is activated in plasmid-free segregants, eventually leading to the elimination of these cells.

U2 - 10.1006/jmbi.1995.0186

DO - 10.1006/jmbi.1995.0186

M3 - Journal article

SN - 0022-2836

VL - 247

SP - 859

EP - 873

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

IS - 5

ER -