Analysis of leaky viral translation termination codons in vivo by transient expression of improved ?-glucuronidase vectors

Skuzeski, James M.; Nichols, Lindy M.; Gesteland, Raymond F.

doi:10.1007/bf00017725

Cited by 92 publications

(78 citation statements)

References 57 publications

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“…All effector genes were placed under the control of the CaMV 35S doubleenhancer promoter and translational enhancer as described (Skuzeski et al, 1990;Tiwari et al, 2001Tiwari et al, , 2003. Effector genes encoding full-length ARF5, GD-5MC, GD-IAA17wt, GD-IAA17mII, and VP16-IAA17mImII have been described previously (Ulmasov et al,1999;Tiwari et al, 2001Tiwari et al, , 2003.…”

Section: Effector Genesmentioning

confidence: 99%

Aux/IAA Proteins Contain a Potent Transcriptional Repression Domain

2004

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Aux/IAA proteins are short-lived nuclear proteins that repress expression of primary/early auxin response genes in protoplast transfection assays. Repression is thought to result from Aux/IAA proteins dimerizing with auxin response factor (ARF) transcriptional activators that reside on auxin-responsive promoter elements, referred to as AuxREs. Most Aux/IAA proteins contain four conserved domains, designated domains I, II, III, and IV. Domain II and domains III and IV play roles in protein stability and dimerization, respectively. A clear function for domain I had not been established. Results reported here indicate that domain I in Aux/IAA proteins is an active repression domain that is transferable and dominant over activation domains. An LxLxL motif within domain I is important for conferring repression. The dominance of Aux/IAA repression domains over activation domains in ARF transcriptional activators provides a plausible explanation for the repression of auxin response genes via ARF-Aux/IAA dimerization on auxin-responsive promoters

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Section: Effector Genesmentioning

confidence: 99%

Aux/IAA Proteins Contain a Potent Transcriptional Repression Domain

2004

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“…IAA4, IAA9 and BDL/IAA12 effector constructs encoding full-length Aux/IAA proteins have been described elsewhere (Tiwari et al, 2001). All effector genes were placed under control of the CaMV 35S double enhancer promoter followed by a translational enhancer from the tobacco mosaic virus 5′ leader (Skuzeski et al, 1990) and contained a 3′ nopaline synthase untranslated region (Ulmasov et al, 1995). Isolation of carrot protoplasts, transfections and GUS activity assays were performed as described previously (Ulmasov et al, 1995;Tiwari et al, 2003).…”

Section: Protoplast Transfection Assaysmentioning

confidence: 99%

Overlapping and non-redundant functions of theArabidopsisauxin response factorsMONOPTEROSandNONPHOTOTROPIC HYPOCOTYL 4

et al. 2004

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Transcription factors of the auxin response factor (ARF) family have been implicated in auxin-dependent gene regulation, but little is known about the functions of individual ARFs in plants. Here, interaction assays, expression studies and combinations of multiple loss- and gain-of-function mutants were used to assess the roles of two ARFs, NONPHOTOTROPIC HYPOCOTYL 4 (NPH4/ARF7)and MONOPTEROS (MP/ARF5), in Arabidopsis development. Both MP and NPH4 interact strongly and selectively with themselves and with each other,and are expressed in vastly overlapping domains. We show that the regulatory properties of both genes are far more related than suggested by their single mutant phenotypes. NPH4 and MP are capable of controlling both axis formation in the embryo and auxin-dependent cell expansion. Interaction of MP and NPH4 in Arabidopsis plants is indicated by their joint requirement in a number of auxin responses and by synergistic effects associated with the co-overexpression of both genes. Finally, we demonstrate antagonistic interaction between ARF and Aux/IAAgene functions in Arabidopsis development. Overexpression of MP suppresses numerous defects associated with a gain-of-function mutation in BODENLOS (BDL)/IAA12. Together these results provide evidence for the biological relevance of ARF-ARF and ARF-Aux/IAA interaction in Arabidopsis plants and demonstrate that an individual ARF can act in both invariantly programmed pattern formation as well as in conditional responses to external signals.

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“…This readthrough process is essential for viability of the virus and controls the level of RNA replicase (Ishikawa et al, 1986). The five nucleotides following the stop codon with the consensus sequence UAG CAR YYA, including the cytosine nucleotide immediately 3h to the UAG stop codon, play an important role in readthrough of the TMV stop coding signal, stimulating 2-5 % suppression (Skuzeski & Atkins, 1990 ;Goelet et al, 1982 ;Skuzeski et al, 1991 ; Table 2). Since no RNA secondary structural elements have been identified in the leaky stop codon environment (Goelet et al, 1982), it seems likely that the 3h stop codon context alone directs readthrough (Skuzeski et al, 1991).…”

Section: Programmed Stop Codon Readthroughmentioning

confidence: 99%

Endless possibilities: translation termination and stop codon recognition

et al. 2001

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OverviewThe process of protein synthesis can be divided into three main phases : initiation, during which the ribosomal subunits join the mRNA and locate the AUG initiator codon ; elongation, during which sense codons are decoded and the bulk of the polypeptide is made ; and termination, during which a stop codon directs the release of the completed polypeptide from the ribosome. Whereas the general principles of sense codon decoding by transfer RNAs are well established, a clear picture of translation termination and stop codon recognition has hitherto been lacking. Recently however, the use of optimized, complex, reconstituted in vitro termination reactions to identify the roles of key termination factors, and the solution of tertiary structures of termination factors, has allowed a reappraisal of termination factor structure and function. This review will describe these recent advances, many of which have resulted from studies of termination in micro-organisms, and in the light of the new information, discuss models for the mechanism of termination and recognition of the stop codon. While the mechanism of peptide chain termination is normally effective, stop codons are not always recognized efficiently, and during the translation of certain viral RNAs can be suppressed or read through, resulting in the expression of additional coding information. How stop codons are reassigned to encode ' sense ' at a given frequency in some RNAs will be reviewed in the context of current understanding of termination and stop codon recognition mechanisms. The translation termination apparatus in eukaryotes and prokaryotesDuring translation termination, a stop codon located in the ribosomal A-site is recognized by a release factor or release factor complex, which binds the ribosome and triggers release of the nascent peptide. In eukaryotes, translation is terminated by a heterodimer consisting of two proteins, release factors eRF1 and eRF3, which interact in vivo (Frolova et al., 1994 ;Zhouravleva et al., 1995 ;Stansfield et al., 1995). eRF1 recognizes all three stop codons and triggers peptidyl-tRNA hydrolysis by the ribosome, releasing the nascent peptide (Frolova et al., 1994 ; Drugeon et al., 1997). Eukaryote termination efficiency is enhanced by the GTPase release factor eRF3, the second component of the heterodimer eRF complex. In response to a stop codon in the ribosomal A-site, formation of a quaternary complex comprising the ribosome, eRF1, GTP and eRF3 triggers GTP hydrolysis and enhances the rate of peptidyl release (Zhouravleva et al., 1995 ; Frolova et al., 1994 ; Fig. 1). In yeast, eRF1 and eRF3 are encoded by essential genes (SUP35 and SUP45, respectively), mutations in which produce nonsense suppression phenotypes (Stansfield & Tuite, 1994).In contrast to eukaryotes, the role of stop codon recognition during translation termination in eubacteria is divided between two so-called class 1 release factors, RF1 and RF2, which in Escherichia coli are encoded by the essential prfA and prfB genes, respectively (Scolni...

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Analysis of leaky viral translation termination codons in vivo by transient expression of improved ?-glucuronidase vectors

Cited by 92 publications

References 57 publications

Aux/IAA Proteins Contain a Potent Transcriptional Repression Domain

Aux/IAA Proteins Contain a Potent Transcriptional Repression Domain

Overlapping and non-redundant functions of theArabidopsisauxin response factorsMONOPTEROSandNONPHOTOTROPIC HYPOCOTYL 4

Endless possibilities: translation termination and stop codon recognition

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