Biophysical characterization of Gir2, a highly acidic protein of Saccharomyces cerevisiae with anomalous electrophoretic behavior

Alves, Viviane S.; Pimenta, Daniel C.; Sattlegger, Evelyn; Castilho, Beatriz A.

doi:10.1016/j.bbrc.2003.12.086

Cited by 41 publications

(35 citation statements)

References 19 publications

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“…Although the function of Gir2 is unknown, the N terminus (aa 5 to 156) shares sequence similarity to the Nterminal GI domain of Gcn2 (34), a polyribosome-associated protein (47). Interestingly, native Gir2 seems to be highly unstructured, perhaps only adopting a folded conformation in the presence of binding partners (1,2).…”

Section: Resultsmentioning

confidence: 99%

Saccharomyces cerevisiae Rbg1 Protein and Its Binding Partner Gir2 Interact on Polyribosomes with Gcn1

et al. 2009

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Rbg1 is a previously uncharacterized protein of Saccharomyces cerevisiae belonging to the Obg/CgtA subfamily of GTP-binding proteins whose members are involved in ribosome function in both prokaryotes and eukaryotes. We show here that Rbg1 specifically associates with translating ribosomes. In addition, in this study proteins were identified that interact with Rbg1 by yeast two-hybrid screening and include Tma46, Ygr250c, Yap1, and Gir2. Gir2 contains a GI (Gcn2 and Impact) domain similar to that of Gcn2, an essential factor of the general amino acid control pathway required for overcoming amino acid shortage. Interestingly, we found that Gir2, like Gcn2, interacts with Gcn1 through its GI domain, and overexpression of Gir2, under conditions mimicking amino acid starvation, resulted in inhibition of growth that could be reversed by Gcn2 co-overexpression. Moreover, we found that Gir2 also cofractionated with polyribosomes, and this fractionation pattern was partially dependent on the presence of Gcn1. Based on these findings, we conclude that Rbg1 and its interacting partner Gir2 associate with ribosomes, and their possible biological roles are discussed.

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Section: Resultsmentioning

confidence: 99%

Saccharomyces cerevisiae Rbg1 Protein and Its Binding Partner Gir2 Interact on Polyribosomes with Gcn1

et al. 2009

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“…The anomalous electrophoretic behavior of Hbr1p may be explained by its high negative charge, which decreases LDS binding (38,43,44). Notably, AD-004 protein is less highly charged and lacks this anomaly (16).…”

Section: Discussionmentioning

confidence: 99%

“…These values change only slightly when the His 6 tag is included in the calculation (Ϫ47 and 4.12, respectively). Highly acidic proteins can exhibit electrophoretic mobilities differing from that expected for a completely unfolded globular protein (38). In most cases, protein mobility is decreased, resulting in an overestimation of the true mass, which may be explained by decreased detergent binding.…”

Section: Hbr1mentioning

confidence: 96%

ATP Binding to Hemoglobin Response Gene 1 Protein Is Necessary for Regulation of the Mating Type Locus in Candida albicans

Peterson

Pendrak

Roberts

2011

Journal of Biological Chemistry

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HBR1 (hemoglobin response gene 1) is an essential gene inCandida albicans that positively regulates mating type locus MTL␣ gene expression and thereby regulates cell type-specific developmental genes. Hbr1p contains a phosphate-binding loop (P-loop), a highly conserved motif characteristic of ATP-and GTP-binding proteins. Recombinant Hbr1p was isolated in an oligomeric state that specifically bound ATP with K d ϳ2 M. ATP but not ADP, AMP, GTP, or dATP specifically protected Hbr1p from proteolysis by trypsin. Site-directed mutagenesis of the highly conserved P-loop lysine (K22Q) and the less conserved glycine (G19S) decreased the binding affinity for soluble ATP and ATP immobilized through its ␥-phosphate. ATP bound somewhat more avidly than ATP␥S to wild type and mutant Hbr1p. Although Hbr1p exhibits sequence motifs characteristic of adenylate kinases, and adenylate kinase and ATPase activities have been reported for the apparent human ortholog of Hbr1p, assays for adenylate kinase activity, autophosphorylation, and ATPase activity proved negative. Overexpression of wild type but not the mutant forms of Hbr1p restored MTl␣2 expression in an HBR1/hbr1 mutant, indicating that ATP binding to the P-loop is necessary for this function of Hbr1p.

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“…One of them, rErum7380, had a molecular weight of 44 kDa which was larger than the expected 34 kDa. SDS-PAGE can yield anomalous mobility for glycoproteins or proteins with low pI (Marciani and Papamatheakis, 1978;Alves et al, 2004). It is not known whether rErum7380 is a glycoprotein but it had a predicted pI value of 4.16 which was the lowest of all the five proteins and this could have affected its mobility.…”

Section: Discussionmentioning

confidence: 99%

In vitro and in vivo evaluation of five low molecular weight proteins of Ehrlichia ruminantium as potential vaccine components

Sebatjane

Pretorius

Liebenberg

et al. 2010

Veterinary Immunology and Immunopathology

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Biophysical characterization of Gir2, a highly acidic protein of Saccharomyces cerevisiae with anomalous electrophoretic behavior

Cited by 41 publications

References 19 publications

Saccharomyces cerevisiae Rbg1 Protein and Its Binding Partner Gir2 Interact on Polyribosomes with Gcn1

Saccharomyces cerevisiae Rbg1 Protein and Its Binding Partner Gir2 Interact on Polyribosomes with Gcn1

ATP Binding to Hemoglobin Response Gene 1 Protein Is Necessary for Regulation of the Mating Type Locus in Candida albicans

In vitro and in vivo evaluation of five low molecular weight proteins of Ehrlichia ruminantium as potential vaccine components

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