The Yeast Peptide Elongation Factor 3 (EF-3) Carries an Active Site for ATP Hydrolysis Which Can Interact with Various Nucleoside Triphosphates in the Absence of Ribosomes1

1988 ◽  
Vol 104 (3) ◽  
pp. 445-450 ◽  
Author(s):  
Masazumi Miyazaki ◽  
Masahiro Uritani ◽  
Hideto Kagiyama
Keyword(s):  
Active Site ◽  
Atp Hydrolysis ◽  
Elongation Factor ◽  
Nucleoside Triphosphates ◽  
Elongation Factor 3 ◽  
Peptide Elongation

scholarly journals Domain and Nucleotide Dependence of the Interaction between Saccharomyces cerevisiae Translation Elongation Factors 3 and 1A

2006 ◽  
Vol 281 (43) ◽  
pp. 32318-32326 ◽  
Author(s):  
Monika Anand ◽  
Bharvi Balar ◽  
Rory Ulloque ◽  
Stephane R. Gross ◽  
Terri Goss Kinzy
Keyword(s):  
Atp Hydrolysis ◽  
Elongation Factor ◽  
Temperature Sensitive ◽  
Translation Elongation ◽  
Eukaryotic Translation ◽  
Domain Mapping ◽  
Elongation Factor 3 ◽  
Domain Iii ◽  
A Site ◽  
Trna Binding

Eukaryotic translation elongation factor 3 (eEF3) is a fungal-specific ATPase proposed to catalyze the release of deacylated-tRNA from the ribosomal E-site. In addition, it has been shown to interact with the aminoacyl-tRNA binding GTPase elongation factor 1A (eEF1A), perhaps linking the E and A sites. Domain mapping demonstrates that amino acids 775–980 contain the eEF1A binding sites. Domain III of eEF1A, which is also involved in actin-related functions, is the site of eEF3 binding. The binding of eEF3 to eEF1A is enhanced by ADP, indicating the interaction is favored post-ATP hydrolysis but is not dependent on the eEF1A-bound nucleotide. A temperature-sensitive P915L mutant in the eEF1A binding site of eEF3 has reduced ATPase activity and affinity for eEF1A. These results support the model that upon ATP hydrolysis, eEF3 interacts with eEF1A to help catalyze the delivery of aminoacyl-tRNA at the A-site of the ribosome. The dynamics of when eEF3 interacts with eEF1A may be part of the signal for transition of the post to pre-translocational ribosomal state in yeast.

Functional Role and Biochemical Properties of Yeast Peptide Elongation Factor 3 (EF-3)

1990 ◽  
pp. 557-566 ◽  
Author(s):  
Masazumi Miyazaki ◽  
Masahiro Uritani ◽  
Yoshihisa Kitaoka ◽  
Kazuko Ogawa ◽  
Hideto Kagiyama
Keyword(s):  
Functional Role ◽  
Elongation Factor ◽  
Biochemical Properties ◽  
Elongation Factor 3 ◽  
Peptide Elongation

scholarly journals Reformulation of an extant ATPase active site to mimic ancestral GTPase activity reveals a nucleotide base requirement for function

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Taylor B Updegrove ◽  
Jailynn Harke ◽  
Vivek Anantharaman ◽  
Jin Yang ◽  
Nikhil Gopalan ◽  
...  
Keyword(s):  
Active Site ◽  
Atp Hydrolysis ◽  
Binding Pocket ◽  
Gtpase Activity ◽  
Biological Functions ◽  
Gtp Hydrolysis ◽  
Nucleotide Base ◽  
Nucleoside Triphosphates ◽  
Nucleotide Specificity ◽  
Hydrolysis Of

Hydrolysis of nucleoside triphosphates releases similar amounts of energy. However, ATP hydrolysis is typically used for energy-intensive reactions, whereas GTP hydrolysis typically functions as a switch. SpoIVA is a bacterial cytoskeletal protein that hydrolyzes ATP to polymerize irreversibly during Bacillus subtilis sporulation. SpoIVA evolved from a TRAFAC class of P-loop GTPases, but the evolutionary pressure that drove this change in nucleotide specificity is unclear. We therefore reengineered the nucleotide-binding pocket of SpoIVA to mimic its ancestral GTPase activity. SpoIVAGTPase functioned properly as a GTPase but failed to polymerize because it did not form an NDP-bound intermediate that we report is required for polymerization. Further, incubation of SpoIVAGTPase with limiting ATP did not promote efficient polymerization. This approach revealed that the nucleotide base, in addition to the energy released from hydrolysis, can be critical in specific biological functions. We also present data suggesting that increased levels of ATP relative to GTP at the end of sporulation was the evolutionary pressure that drove the change in nucleotide preference in SpoIVA.

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