scholarly journals Characterization of Substitution Mutations of eIF4G Gene Generated through Adenine Base Editors in Rice

2021 ◽  
Vol 108 (september) ◽  
Author(s):  
Yaiphabi Kumam ◽  
◽  
Rajadurai C ◽  
Kumar K K ◽  
Varanavasiappan S ◽  
...  
Keyword(s):  
Rice Genome ◽  
Rice Cultivar ◽  
Translation Initiation Factor ◽  
Initiation Factor ◽  
Base Substitution ◽  
Amino Acid Residues ◽  
Substitution Mutations ◽  
Mutation Efficiency ◽  
Adenine Base

Adenine base editor (ABE) creates A to G transitions within its editing window. In the present study, an ABE was used to target a stretch of six amino acid residues, VLFPNL in translation initiation factor four gamma (eIF4G) gene of rice. Agrobacterium-mediated transformation of rice cultivar ASD16 resulted in T0 events with high mutation efficiency of 89.29 %. Substitution mutations of A > G occurred within the editing window of four to eight bases at A7 > G7 (74.67 %) and A4 > G4 (2.46 %). Non-canonical substitutions of G > C/A was also observed at G15 > C15 (9.29 %) and G8 > A8 (1.15 %). A total of 15 missense base substitution events affecting the target residue was identified. Taken together, the present study showed that ABEs create unexpected base substitutions besides efficient canonical editing of A > G in the rice genome

scholarly journals Adenine Base Editor Creates Novel Substitution Mutations in eIF4G Gene of Rice

2021 ◽  
Vol 108 (special) ◽  
Author(s):  
Yaiphabi Kumam ◽  
◽  
Rajadurai G ◽  
Kumar K K ◽  
Varanavasiappan S ◽  
...  
Keyword(s):  
Translation Initiation Factor ◽  
Initiation Factor ◽  
Missense Mutations ◽  
Amino Acid Residues ◽  
Single Nucleotide ◽  
Base Editing ◽  
Rice Tungro Disease ◽  
Substitution Mutations ◽  
Resistant Genotypes ◽  
Adenine Base

Two single nucleotide polymorphic mutations and deletion affecting Y1059V1060V1061 amino acid residues in a host translation initiation factor four gamma (eIF4G) gene in rice are reported to confer resistance to rice tungrospherical virus in resistant genotypes. A CRISPR-based adenine base editing vector was used to target these residues in a susceptible indica cultivar, ASD16.Agrobacterium-mediated transformation of ASD16 generated 16 missense mutants and two deletion mutants. Substitution mutations occurred at A5> G5 and A4> G4, where 5.5 % and 3.37 % of adenosines got converted to guanosines, respectively. The mutantsgenerated had missense mutations affecting the YVV residues and the residues immediately adjacent to YVV.Thus,these novel mutationsare promising candidates in imparting resistance against rice tungro disease.

scholarly journals Reaction of lettuce genotypes to Lettuce mosaic virus-Most (LMV-Most) and characterization of the translation factor eIF4E

2018 ◽  
Vol 53 (1) ◽  
pp. 125-129
Author(s):  
Mônika Fecury Moura ◽  
Norberto da Silva ◽  
Maria Isabel Motta Hoffmann ◽  
Marcelo Agenor Pavan ◽  
Renate Krause-Sakate
Keyword(s):  
Mosaic Virus ◽  
Translation Initiation Factor ◽  
Initiation Factor ◽  
Coding Region ◽  
Rt Pcr ◽  
Lettuce Mosaic Virus ◽  
Eukaryotic Translation ◽  
Translation Factor ◽  
Eukaryotic Translation Initiation

Abstract: The objective of this work was to evaluate lettuce genotypes for their reaction to Lettuce mosaic virus (LMV; Most-type, isolate AF-199) and variations of the eukaryotic translation initiation factor eIF4E. All inoculated genotypes were susceptible to LMV, which was detected by RT-PCR using specific primer pairs. However, the accessions 169501, 169501C, 172918A, and 162499 showed late development of symptoms that appeared only on the inoculated leaves. Sequencing of the coding region of eIF4E showed that these genotypes have an eIF4E0 (mol 0 ) standard typical for their susceptibility to LMV, indicating that the phenotype found is not correlated to nucleotide variations in this translation factor.

scholarly journals Mutational Analysis of Plant Cap-Binding Protein eIF4E Reveals Key Amino Acids Involved in Biochemical Functions and Potyvirus Infection

2008 ◽  
Vol 82 (15) ◽  
pp. 7601-7612 ◽  
Author(s):  
Sylvie German-Retana ◽  
Jocelyne Walter ◽  
Bénédicte Doublet ◽  
Geneviève Roudet-Tavert ◽  
Valérie Nicaise ◽  
...  
Keyword(s):  
Amino Acids ◽  
Mutational Analysis ◽  
Binding Protein ◽  
Point Mutations ◽  
Mrna Translation ◽  
Translation Initiation Factor ◽  
Natural Resistance ◽  
Initiation Factor ◽  
Amino Acid Residues ◽  
Eukaryotic Translation

ABSTRACT The eukaryotic translation initiation factor 4E (eIF4E) (the cap-binding protein) is involved in natural resistance against several potyviruses in plants. In lettuce, the recessive resistance genes mo1 1 and mo1 2 against Lettuce mosaic virus (LMV) are alleles coding for forms of eIF4E unable, or less effective, to support virus accumulation. A recombinant LMV expressing the eIF4E of a susceptible lettuce variety from its genome was able to produce symptoms in mo1 1 or mo1 2 varieties. In order to identify the eIF4E amino acid residues necessary for viral infection, we constructed recombinant LMV expressing eIF4E with point mutations affecting various amino acids and compared the abilities of these eIF4E mutants to complement LMV infection in resistant plants. Three types of mutations were produced in order to affect different biochemical functions of eIF4E: cap binding, eIF4G binding, and putative interaction with other virus or host proteins. Several mutations severely reduced the ability of eIF4E to complement LMV accumulation in a resistant host and impeded essential eIF4E functions in yeast. However, the ability of eIF4E to bind a cap analogue or to fully interact with eIF4G appeared unlinked to LMV infection. In addition to providing a functional mutational map of a plant eIF4E, this suggests that the role of eIF4E in the LMV cycle might be distinct from its physiological function in cellular mRNA translation.

scholarly journals A Novel Function of the MA-3 Domains in Transformation and Translation Suppressor Pdcd4 Is Essential for Its Binding to Eukaryotic Translation Initiation Factor 4A

2004 ◽  
Vol 24 (9) ◽  
pp. 3894-3906 ◽  
Author(s):  
Hsin-Sheng Yang ◽  
Myung-Haing Cho ◽  
Halina Zakowicz ◽  
Glenn Hegamyer ◽  
Nahum Sonenberg ◽  
...  
Keyword(s):  
Amino Acid ◽  
Translation Initiation ◽  
Binding Activity ◽  
Translation Initiation Factor ◽  
Initiation Factor ◽  
Amino Acid Residues ◽  
Eukaryotic Translation Initiation Factor ◽  
Stem Loop ◽  
Eukaryotic Translation ◽  
Eukaryotic Translation Initiation

ABSTRACT Αn α-helical MA-3 domain appears in several translation initiation factors, including human eukaryotic translation initiation factor 4G (eIF4G) and DAP-5/NAT1/p97, as well as in the tumor suppressor Pdcd4. The function of the MA-3 domain is, however, unknown. C-terminal eIF4G (eIG4Gc) contains an MA-3 domain that is located within the eIF4A-binding region, suggesting a role for eIF4A binding. Interestingly, C-terminal DAP-5/NAT1/p97 contains an MA-3 domain, but it does not bind to eIF4A. Mutation of amino acid residues conserved between Pdcd4 and eIF4Gc but not in DAP-5/NAT1/p97 to the amino acid residues found in the DAP-5/NAT1/p97 indicates that some of these amino acid residues within the MA-3 domain are critical for eIF4A-binding activity. Six Pdcd4 mutants (Pdcd4E249K, Pdcd4D253A, Pdcd4D414K, Pdcd4D418A, Pdcd4E249K,D414K, and Pdcd4D253A,D418A) lost >90% eIF4A-binding activity. Mutation of the corresponding amino acid residues in the eIF4Gc also produced similar results, as seen for Pdcd4. These results demonstrate that the MA-3 domain is important for eIF4A binding and explain the ability of Pdcd4 or eIF4Gc but not DAP-5/NAT1/p97 to bind to eIF4A. Competition experiments indicate that Pdcd4 prevents ca. 60 to 70% of eIF4A binding to eIF4Gc at a Pdcd4/eIF4A ratio of 1:1, but mutants Pdcd4D253A and Pdcd4D253A,D418A do not. Translation of stem-loop structured mRNA is susceptible to inhibition by wild-type Pdcd4 but not by Pdcd4D253A, Pdcd4D418A, or Pdcd4D235A,D418A. Together, these results indicate that not only binding to eIF4A but also prevention of eIF4A binding to the MA-3 domain of eIF4Gc contributes to the mechanism by which Pdcd4 inhibits translation.

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