scholarly journals Interactions between RNase P protein subunits in Archaea

Archaea ◽  
2004 ◽  
Vol 1 (4) ◽  
pp. 247-254 ◽  
Author(s):  
Thomas A. Hall ◽  
James W. Brown
Keyword(s):  
Protein Interactions ◽  
Rnase P ◽  
Ribonuclease P ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Protein Subunits ◽  
P Protein ◽  
Yeast Two Hybrid System ◽  
Methanothermobacter Thermoautotrophicus ◽  
Two Hybrid

A yeast two-hybrid system was used to identify protein–protein interactions between the ribonuclease P (RNase P) protein subunits Mth11p, Mth687p, Mth688p and Mth1618p from the archaeonMethanothermobacter thermoautotrophicus. Clear interactions between Mth688p and Mth687p, and between Mth1618p and Mth11p, were confirmed byHIS3andLacZreporter expression. Weaker interactions of Mth687p and Mth688p with Mth11p, and Mth11p with itself, are also suggested. These interactions resemble, and confirm, those previously seen among the homologs of these proteins in the more complex yeast RNase P holoenzyme.

Determination of Protein-Protein Interactions of ICln by the Yeast Two-Hybrid System

2001 ◽  
Vol 11 (1) ◽  
pp. 55-60 ◽  
Author(s):  
Andreas Schmarda ◽  
Friedrich Fresser ◽  
Martin Gschwentner ◽  
Johannes Fürst ◽  
Markus Ritter ◽  
...  
Keyword(s):  
Hybrid System ◽  
Protein Interactions ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Yeast Two Hybrid System ◽  
Two Hybrid

scholarly journals Upf1p, Nmd2p, and Upf3p are interacting components of the yeast nonsense-mediated mRNA decay pathway.

1997 ◽  
Vol 17 (3) ◽  
pp. 1580-1594 ◽  
Author(s):  
F He ◽  
A H Brown ◽  
A Jacobson
Keyword(s):  
Protein Interactions ◽  
Mrna Decay ◽  
Dominant Negative ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Yeast Two Hybrid System ◽  
Essential Components ◽  
Nonsense Mediated Mrna Decay ◽  
Two Hybrid ◽  
Interacting Components

Rapid turnover of nonsense-containing mRNAs in Saccharomyces cerevisiae is dependent on Upf1p, Nmd2p, and Upf3p, the products of the UPF1, NMD2/UPF2, and UPF3 genes, respectively. We showed previously that Upf1p and Nmd2p interact and that this interaction is required for nonsense-mediated mRNA decay (F. He and A. Jacobson, Genes Dev. 9:437-454, 1995; F. He, A. H. Brown, and A. Jacobson, RNA 2:153-170, 1996). In this study we have used the yeast two-hybrid system to define other protein-protein interactions among the essential components of this decay pathway. Nmd2p-Upf3p and Upf1p-Upf3p interactions were identified, and the respective domains involved in these interactions were delineated by deletion analysis. The domains of Upf1p and Upf3p putatively involved in their mutual interaction were found to correspond to the domains on the two proteins which interact with Nmd2p, suggesting that Nmd2p bridges Upf1p and Upf3p. This conclusion was reinforced by experiments showing that: (i) deletion of NMD2 completely abolishes interactions between Upf1p and Upf3p and (ii) overexpression of full-length Nmd2p or Nmd2p fragments that retain Upf1p- and Upf3p-interacting domains promotes 10- to 200-fold enhancement of Upf1p-Nmd2p-Upf3p complex formation. These results; the observation that cells harboring either single or multiple deletions of UPF1, NMD2, and UPF3 inhibit nonsense-mediated mRNA decay to the same extent; and an analysis of the possible targets of a dominant-negative NMD2 allele indicate that Upf1p, Nmd2p, Upf3p, and at least one other factor are functionally dependent, interacting components of the yeast nonsense-mediated mRNA decay pathway.

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