Molecular docking uncovers TSPY binds more efficiently with eEF1A2 compared to eEF1A1

Abstract

Testis-specific protein, Y-encoded (TSPY) binds to eukaryotic translation elongation factor 1 alpha (eEF1A) at its SET/NAP domain that is essential for the elongation during protein synthesis implicated with normal spermatogenesis. The eEF1A exists in two forms, eEF1A1 (alpha 1) and eEF1A2 (alpha 2), encoded by separate loci. Despite critical interplay of the TSPY and eEF1A proteins, literature remained silent on the residues playing significant roles during such interactions. We deduced 3D structures of TSPY and eEF1A variants by comparative modeling (Modeller 9.13) and assessed protein-protein interactions employing HADDOCK docking. Pairwise alignment using EMBOSS Needle for eEF1A1 and eEF1A2 proteins revealed high degree (~92%) of homology. Efficient binding of TSPY with eEF1A2 as compared to eEF1A1 was observed, in spite of the occurrence of significant structural similarities between the two variants. We also detected strong interactions of domain III followed by domains II and I of both eEF1A variants with TSPY. In the process, seven interacting residues of TSPY's NAP domain namely, Asp 175, Glu 176, Asp 179, Tyr 183, Asp 240, Glu 244, and Tyr 246 common to both eEF1A variants were detected. Additionally, six lysine residues observed in eEF1A2 suggest their possible role in TSPY-eEF1A2 complex formation essential for germ cell development and spermatogenesis. Thus, more efficient binding of TSPY with eEF1A2 as compared to that of eEF1A1 established autonomous functioning of these two variants. Studies on mutated protein following similar approach would uncover the causative obstruction, between the interacting partners leading to deeper understanding on the structure-function relationship.

Comparison of Y-chromosome-linked TSPY, TSPY2, and PRAMEY genes in Taurus cattle, yaks, and interspecific hybrid bulls

“…TSPY is thought to play an important role in male spermatogenesis and might be a potential risk factor for male infertility (Krausz et al, 2010). Human TSPY binds to cell cycle controlling cyclins B1 (Li and Lau, 2008), and translational elongation factors such as eEF1A1 and eEF1A2 (Kido and Lau, 2008;Panwar et al, 2015). Recently, it has been reported human TSPY suppresses USP7-mediated p53 function and promotes spermatogonial proliferation (Shen et al, 2018).…”

Spermatogenesis-associated proteins at different developmental stages of buffalo testicular seminiferous tubules identified by comparative proteomic analysis

“…Eukaryotic elongation factor 1 is a translational multimolecular complex reported in higher eukaryotes and the universal regulator of mitotic phase . EEF1A1 is essential for the elongation during protein synthesis associated with normal spermatogenesis , and has even been found to be a target of an antispermatogenic compound . Also, EEF1G, a member of eukaryotic elongation factor 1, is an important component of the HIV‐1 reverse transcription complex .…”

Nudix‐type RNA pyrophosphohydrolase provides homeostasis of virulence factor pyocyanin and functions as a global regulator in Pseudomonas aeruginosa

“… Alignment of P. aeruginosa PA0336 ( Pa RppH) and E. coli RppH ( Ec RppH) amino acid sequences. The alignment was performed using Needle global alignment tool (Panwar et al ., ). Identical amino acids are boxed in black, and similar ones are in grey.…”

“…The cellular level of rpsT P1 transcript was determined by quantitative RT-PCR in the DrppH mutant and the DrppH mutant complemented by pPA.2 construct, pEC.1 plasmid carrying the wildtype allele of the E. coli rppH gene, pPA.2 M plasmid carrying the mutated allele of PA0336 or pPA.3 vector carrying Pseudomonas nudC gene encoding NudC Nudix hydrolase active on NADH (Table 1 and 2) (Modzelan et al, 2014). The Pseudomonas PA0336 hydrolase The alignment was performed using Needle global alignment tool (Panwar et al, 2009). Identical amino acids are boxed in black, and similar ones are in grey.…”