Identification of amino acid residues in protein SRP72 required for binding to a kinked 5e motif of the human signal recognition particle RNA

<p>Abstract</p> <p>Background</p> <p>Human cells depend critically on the signal recognition particle (SRP) for the sorting and delivery of their proteins. The SRP is a ribonucleoprotein complex which binds to signal sequences of secretory polypeptides as they emerge from the ribosome. Among the six proteins of the eukaryotic SRP, the largest protein, SRP72, is essential for protein targeting and possesses a poorly characterized RNA binding domain.</p> <p>Results</p> <p>We delineated the minimal region of SRP72 capable of forming a stable complex with an SRP RNA fragment. The region encompassed residues 545 to 585 of the full-length human SRP72 and contained a lysine-rich cluster (KKKKKKKKGK) at postions 552 to 561 as well as a conserved Pfam motif with the sequence PDPXRWLPXXER at positions 572 to 583. We demonstrated by site-directed mutagenesis that both regions participated in the formation of a complex with the RNA. In agreement with biochemical data and results from chymotryptic digestion experiments, molecular modeling of SRP72 implied that the invariant W577 was located inside the predicted structure of an RNA binding domain. The 11-nucleotide 5e motif contained within the SRP RNA fragment was shown by comparative electrophoresis on native polyacrylamide gels to conform to an RNA kink-turn. The model of the complex suggested that the conserved A240 of the K-turn, previously identified as being essential for the binding to SRP72, could protrude into a groove of the SRP72 RNA binding domain, similar but not identical to how other K-turn recognizing proteins interact with RNA.</p> <p>Conclusions</p> <p>The results from the presented experiments provided insights into the molecular details of a functionally important and structurally interesting RNA-protein interaction. A model for how a ligand binding pocket of SRP72 can accommodate a new RNA K-turn in the 5e region of the eukaryotic SRP RNA is proposed.</p

On the Shoulders of Giants: The Growing Impact of Older Articles

In this paper, we examine the evolution of the impact of older scholarly articles. We attempt to answer four questions. First, how often are older articles cited and how has this changed over time. Second, how does the impact of older articles vary across different research fields. Third, is the change in the impact of older articles accelerating or slowing down. Fourth, are these trends different for much older articles. To answer these questions, we studied citations from articles published in 1990-2013. We computed the fraction of citations to older articles from articles published each year as the measure of impact. We considered articles that were published at least 10 years before the citing article as older articles. We computed these numbers for 261 subject categories and 9 broad areas of research. Finally, we repeated the computation for two other definitions of older articles, 15 years and older and 20 years and older. There are three conclusions from our study. First, the impact of older articles has grown substantially over 1990-2013. In 2013, 36% of citations were to articles that are at least 10 years old; this fraction has grown 28% since 1990. The fraction of older citations increased over 1990-2013 for 7 out of 9 broad areas and 231 out of 261 subject categories. Second, the increase over the second half (2002-2013) was double the increase in the first half (1990-2001). Third, the trend of a growing impact of older articles also holds for even older articles. In 2013, 21% of citations were to articles >= 15 years old with an increase of 30% since 1990 and 13% of citations were to articles >= 20 years old with an increase of 36%. Now that finding and reading relevant older articles is about as easy as finding and reading recently published articles, significant advances aren't getting lost on the shelves and are influencing work worldwide for years after