Roles of the N- and C-Terminal Domains of Mammalian Mitochondrial Initiation Factor 3 in Protein Biosynthesis

Bacterial initiation factor 3 (IF3) is organized into N-domain and C-domains separated by a linker. Mitochondrial IF3 (IF3mt) has a similar domain organization although both domains have extensions not found in the bacterial factors. Constructs of the N- and C-domains of IF3mt with and without the connecting linker were prepared. The Kd’s for the binding of full length IF3mt and its C-domain with and without the linker to mitochondrial 28S subunits are 30, 60 and 95 nM respectively, indicating that much of the ribosome binding interactions are mediated by the C-domain. However, the N-domain binds to 28S subunits with only a 10-fold lower affinity than full length IF3mt. This observation indicates that the N-domain of IF3mt has significant contacts with the protein-rich small subunit of mammalian mitochondrial ribosomes. The linker also plays a role in modulating the interactions between the 28S subunit and the factor; it is not just a physical connector between the two domains. The presence of the two domains and the linker may optimize the overall affinity of IF3mt for the ribosome. These results are in sharp contrast to observations with E. coli IF3. Removal of the N-domain drastically reduces the activity of IF3mt in the dissociation of mitochondrial 55S ribosomes although the C-domain itself retains some activity. This residual activity depends significantly on the linker region. The N-domain alone has no effect on the dissociation of ribosomes. Full-length IF3mt reduces the binding of fMet-tRNA to the 28S subunit in the absence of mRNA. Both the C-terminal extension and the linker are required for this effect. IF3mt promotes the formation of a binary complex between IF2mt and fMet-tRNA that may play an important role in mitochondrial protein synthesis. Both domains play a role promoting the formation of this comple

The interaction of mammalian mitochondrial translational initiation factor 3 with ribosomes: evolution of terminal extensions in IF3 mt

Mammalian mitochondrial initiation factor 3 (IF3mt) has a central region with homology to bacterial IF3. This homology region is preceded by an N-terminal extension and followed by a C-terminal extension. The role of these extensions on the binding of IF3mt to mitochondrial small ribosomal subunits (28S) was studied using derivatives in which the extensions had been deleted. The Kd for the binding of IF3mt to 28S subunits is ∼30 nM. Removal of either the N- or C-terminal extension has almost no effect on this value. IF3mt has very weak interactions with the large subunit of the mitochondrial ribosome (39S) (Kd = 1.5 μM). However, deletion of the extensions results in derivatives with significant affinity for 39S subunits (Kd = 0.12−0.25 μM). IF3mt does not bind 55S monosomes, while the deletion derivative binds slightly to these particles. IF3mt is very effective in dissociating 55S ribosomes. Removal of the N-terminal extension has little effect on this activity. However, removal of the C-terminal extension leads to a complex dissociation pattern due to the high affinity of this derivative for 39S subunits. These data suggest that the extensions have evolved to ensure the proper dissociation of IF3mt from the 28S subunits upon 39S subunit joining

The effect of spermine on the initiation of mitochondrial protein synthesis

Polyamines are important in both prokaryotic and eukaryotic translational systems. Spermine is a quaternary aliphatic amine that is cationic under physiological conditions. In this paper, we demonstrate that spermine stimulates fMet-tRNA binding to mammalian mitochondrial 55S ribosomes. The stimulatory effect of spermine is independent of the identity of the mRNA. The degree of stimulation of spermine is the same at all concentrations of mitochondrial initiation factor 2 (IF2mt) and mitochondrial initiation factor 3 (IF3mt). This observation indicates that IF2mt and IF3mt, while essential for initiation, are not the primary components of the translation initiation system affected by spermine. IF3mt dissociates 55S ribosomes detectably in the absence of spermine, but this effect is strongly inhibited in the presence of spermine. This observation indicates that the positive effect of spermine on initiation is not due to an increase in the availability of the small subunits for initiation. Spermine also promotes fMet-tRNA binding to small subunits of the mitochondrial ribosome in the presence of IF2mt. The major effect of spermine in promoting initiation complex formation thus appears to be on the interaction of fMet-tRNA with the ribosome

Book Cover Synthesis from the Summary

The cover is the face of a book and is a point of attraction for the readers. Designing book covers is an essential task in the publishing industry. One of the main challenges in creating a book cover is representing the theme of the book's content in a single image. In this research, we explore ways to produce a book cover using artificial intelligence based on the fact that there exists a relationship between the summary of the book and its cover. Our key motivation is the application of text-to-image synthesis methods to generate images from given text or captions. We explore several existing text-to-image conversion techniques for this purpose and propose an approach to exploit these frameworks for producing book covers from provided summaries. We construct a dataset of English books that contains a large number of samples of summaries of existing books and their cover images. In this paper, we describe our approach to collecting, organizing, and pre-processing the dataset to use it for training models. We apply different text-to-image synthesis techniques to generate book covers from the summary and exhibit the results in this paper.Comment: Accepted as a full paper in AICCSA2022 (19th ACS/IEEE International Conference on Computer Systems and Applications

Interaction of α-synuclein with vesicles that mimic mitochondrial membranes

Abstractα-Synuclein, an intrinsically-disordered protein associated with Parkinson's disease, interacts with mitochondria, but the details of this interaction are unknown. We probed the interaction of α-synuclein and its A30P variant with lipid vesicles by using fluorescence anisotropy and 19F nuclear magnetic resonance. Both proteins interact strongly with large unilamellar vesicles whose composition is similar to that of the inner mitochondrial membrane, which contains cardiolipin. However, the proteins have no affinity for vesicles mimicking the outer mitochondrial membrane, which lacks cardiolipin. The 19F data show that the interaction involves α-synuclein's N-terminal region. These data indicate that the middle of the N-terminal region, which contains the KAKEGVVAAAE repeats, is involved in binding, probably via electrostatic interactions between the lysines and cardiolipin. We also found that the strength of α-synuclein binding depends on the nature of the cardiolipin acyl side chains. Eliminating one double bond increases affinity, while complete saturation dramatically decreases affinity. Increasing the temperature increases the binding of wild-type, but not the A30P variant. The data are interpreted in terms of the properties of the protein, cardiolipin demixing within the vesicles upon binding of α-synuclein, and packing density. The results advance our understanding of α-synuclein's interaction with mitochondrial membranes

The effect of mutated mitochondrial ribosomal proteins S16 and S22 on the assembly of the small and large ribosomal subunits in human mitochondria

Mutations in mitochondrial small subunit ribosomal proteins MRPS16 or MRPS22 cause severe, fatal respiratory chain dysfunction due to impaired translation of mitochondrial mRNAs. The loss of either MRPS16 or MRPS22 was accompanied by the loss of most of another small subunit protein MRPS11. However, MRPS2 was reduced only about 2-fold in patient fibroblasts. This observation suggests that the small ribosomal subunit is only partially able to assemble in these patients. Two large subunit ribosomal proteins, MRPL13 and MRPL15, were present in substantial amounts suggesting that the large ribosomal subunit is still present despite a nonfunctional small subunit