scholarly journals N1-Propargylguanosine Modified mRNA Cap Analogs: Synthesis, Reactivity, and Applications to the Study of Cap-Binding Proteins

Molecules ◽  
2019 ◽  
Vol 24 (10) ◽  
pp. 1899
Author(s):  
Michal Kopcial ◽  
Blazej A. Wojtczak ◽  
Renata Kasprzyk ◽  
Joanna Kowalska ◽  
Jacek Jemielity
Keyword(s):  
Protein Interactions ◽  
Binding Proteins ◽  
Molecular Probes ◽  
Translation Initiation Factor ◽  
Initiation Factor ◽  
Binding Assays ◽  
Fluorescent Properties ◽  
Eukaryotic Translation ◽  
Eukaryotic Translation Initiation ◽  
Fluorescent Tags

The mRNA 5′ cap consists of N7-methylguanosine bound by a 5′,5′-triphosphate bridge to the first nucleotide of the transcript. The cap interacts with various specific proteins and participates in all key mRNA-related processes, which may be of therapeutic relevance. There is a growing demand for new biophysical and biochemical methods to study cap–protein interactions and identify the factors which inhibit them. The development of such methods can be aided by the use of properly designed fluorescent molecular probes. Herein, we synthesized a new class of m7Gp3G cap derivatives modified with an alkyne handle at the N1-position of guanosine and, using alkyne-azide cycloaddition, we functionalized them with fluorescent tags to obtain potential probes. The cap derivatives and probes were evaluated in the context of two cap-binding proteins, eukaryotic translation initiation factor (eIF4E) and decapping scavenger (DcpS). Biochemical and biophysical studies revealed that N1-propargyl moiety did not significantly disturb cap–protein interaction. The fluorescent properties of the probes turned out to be in line with microscale thermophoresis (MST)-based binding assays.

Abstract 321: Deletion Of The Translational Repressors Eukaryotic Translation Initiation Factor 4e Binding Proteins 1 And 2 Protects Against Pressure Overload Induced Heart Failure

2013 ◽  
Vol 113 (suppl_1) ◽  
Author(s):  
zhongbing lu ◽  
Xinli Hu ◽  
Yimin Huang ◽  
Xin Xu ◽  
Ping zhang ◽  
...  
Keyword(s):  
Translation Initiation ◽  
Translational Control ◽  
Binding Proteins ◽  
Pressure Overload ◽  
Left Ventricular ◽  
Translation Initiation Factor ◽  
Initiation Factor ◽  
Double Knockout ◽  
Eukaryotic Translation ◽  
Eukaryotic Translation Initiation

Assembly of the translation initiation machinery is negatively regulated by the eukaryotic translation initiation factor 4E binding proteins, which sequester the mRNA cap-binding protein eIF4E, thus preventing assembly of an intact initiation complex. However, the role of translational control on the development of congestive heart failure (CHF) has not been systematically examined. Here we perturbed translational control in mice by knockout of both 4E binding protein 1 (Eif4ebp1) and 2 (Eif4ebp2) (designated as Eif4ebp1/2 double knockout) to study its impact on left ventricular hypertrophy and CHF resulting from transverse aortic constriction. Eif4ebp1/2 double knockout caused a modest increase in left ventricular mass under basal conditions. However, following transverse aortic constriction, Eif4ebp1/2 double knockout profoundly attenuated the development of CHF and its attendant mortality. Examination of candidate genes involved in the mechanism revealed increased expression of transcription factors for genes governing energy metabolism and mitochondrial biogenesis with corresponding increases in the expression of their target genes. Our data indicate that removing physiological restraints on translation initiation exerts a profound cardiac protective effect against pressure overload induced CHF, suggesting that method(s) to disrupt the function of the 4E binding proteins may be a novel therapeutic approach for preventing or treating CHF.

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