Mapping the role of hRNase P individual protein subunits via CRISPR/Cas9

2020 ◽  
Author(s):  
Constantinos Stathopoulos
Keyword(s):  
Protein Subunits ◽  
Individual Protein

Heterogeneous protein co-assemblies with tunable functional domain stoichiometry

2021 ◽  
Author(s):  
Shaheen Farhadi ◽  
Antonietta Restuccia ◽  
Anthony M Sorrentino ◽  
Andres Cruz-Sanchez ◽  
Gregory A Hudalla
Keyword(s):  
Protein Domains ◽  
Functional Domain ◽  
Protein Subunits ◽  
Individual Protein ◽  
Complex Functions ◽  
The Individual

In nature, the precise heterogeneous co-assembly of different protein domains gives rise to supramolecular machines that perform complex functions through the co-integrated activity of the individual protein subunits. A synthetic...

scholarly journals Multi-omics analysis of glucose-mediated signaling by a moonlighting Gβ protein Asc1/RACK1

PLoS Genetics ◽  
2021 ◽  
Vol 17 (7) ◽  
pp. e1009640
Author(s):  
Shuang Li ◽  
Yuanyuan Li ◽  
Blake R. Rushing ◽  
Sarah E. Harris ◽  
Susan L. McRitchie ◽  
...  
Keyword(s):  
G Protein ◽  
Glucose Sensing ◽  
Heterotrimeric G Proteins ◽  
Glucose Addition ◽  
Protein Subunits ◽  
Individual Gene ◽  
Gene Transcripts ◽  
G Protein Coupled

Heterotrimeric G proteins were originally discovered through efforts to understand the effects of hormones, such as glucagon and epinephrine, on glucose metabolism. On the other hand, many cellular metabolites, including glucose, serve as ligands for G protein-coupled receptors. Here we investigate the consequences of glucose-mediated receptor signaling, and in particular the role of a Gα subunit Gpa2 and a non-canonical Gβ subunit, known as Asc1 in yeast and RACK1 in animals. Asc1/RACK1 is of particular interest because it has multiple, seemingly unrelated, functions in the cell. The existence of such “moonlighting” operations has complicated the determination of phenotype from genotype. Through a comparative analysis of individual gene deletion mutants, and by integrating transcriptomics and metabolomics measurements, we have determined the relative contributions of the Gα and Gβ protein subunits to glucose-initiated processes in yeast. We determined that Gpa2 is primarily involved in regulating carbohydrate metabolism while Asc1 is primarily involved in amino acid metabolism. Both proteins are involved in regulating purine metabolism. Of the two subunits, Gpa2 regulates a greater number of gene transcripts and was particularly important in determining the amplitude of response to glucose addition. We conclude that the two G protein subunits regulate distinct but complementary processes downstream of the glucose-sensing receptor, as well as processes that lead ultimately to changes in cell growth and metabolism.

scholarly journals Repositioning the Sm-Binding Site in Saccharomyces cerevisiae Telomerase RNA Reveals RNP Organizational Flexibility and Sm-Directed 3′-End Formation

2020 ◽  
Vol 6 (1) ◽  
pp. 9 ◽  
Author(s):  
Evan P. Hass ◽  
David C. Zappulla
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Binding Site ◽  
Telomerase Rna ◽  
Telomeric Dna ◽  
Protein Subunits ◽  
Yeast Telomerase ◽  
Rna Biogenesis ◽  
Native Site ◽  
Processing Mechanism

Telomerase RNA contains a template for synthesizing telomeric DNA and has been proposed to act as a flexible scaffold for holoenzyme protein subunits in the RNP. In Saccharomyces cerevisiae, the telomerase RNA, TLC1, is bound by the Sm7 protein complex, which is required for stabilization of the predominant, non-polyadenylated (poly(A)–) TLC1 isoform. However, it remains unclear (1) whether Sm7 retains this function when its binding site is repositioned within TLC1, as has been shown for other TLC1-binding telomerase subunits, and (2) how Sm7 stabilizes poly(A)– TLC1. Here, we first show that Sm7 can stabilize poly(A)– TLC1 even when its binding site is repositioned via circular permutation to several different positions within TLC1, further supporting the conclusion that the telomerase holoenzyme is organizationally flexible. Next, we show that when an Sm site is inserted 5′ of its native position and the native site is mutated, Sm7 stabilizes shorter forms of poly(A)– TLC1 in a manner corresponding to how far upstream the new site was inserted, providing strong evidence that Sm7 binding to TLC1 controls where the mature poly(A)– 3′ is formed by directing a 3′-to-5′ processing mechanism. In summary, our results show that Sm7 and the 3′ end of yeast telomerase RNA comprise an organizationally flexible module within the telomerase RNP and provide insights into the mechanistic role of Sm7 in telomerase RNA biogenesis.

scholarly journals Eye evolution: common use and independent recruitment of genetic components

2009 ◽  
Vol 364 (1531) ◽  
pp. 2819-2832 ◽  
Author(s):  
Pavel Vopalensky ◽  
Zbynek Kozmik
Keyword(s):  
Spatial Organization ◽  
Photoreceptor Cell ◽  
Pigment Cell ◽  
Regulatory Relationship ◽  
Protein Subunits ◽  
Eye Evolution ◽  
Genetic Components ◽  
Organization Analysis ◽  
Animal Phyla

Animal eyes can vary in complexity ranging from a single photoreceptor cell shaded by a pigment cell to elaborate arrays of these basic units, which allow image formation in compound eyes of insects or camera-type eyes of vertebrates. The evolution of the eye requires involvement of several distinct components—photoreceptors, screening pigment and genes orchestrating their proper temporal and spatial organization. Analysis of particular genetic and biochemical components shows that many evolutionary processes have participated in eye evolution. Multiple examples of co-option of crystallins, Gα protein subunits and screening pigments contrast with the conserved role of opsins and a set of transcription factors governing eye development in distantly related animal phyla. The direct regulation of essential photoreceptor genes by these factors suggests that this regulatory relationship might have been already established in the ancestral photoreceptor cell.

scholarly journals The role of individual protein kinase C isoforms in mouse mast cell function and their targeting by the immunomodulatory parasitic worm product, ES-62

2015 ◽  
Vol 168 (1) ◽  
pp. 31-40 ◽  
Author(s):  
Kara S. Bell ◽  
Lamyaa Al-Riyami ◽  
Felicity E. Lumb ◽  
Graham J. Britton ◽  
Alastair W. Poole ◽  
...  
Keyword(s):  
Mast Cell ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
Cell Function ◽  
Individual Protein ◽  
Kinase C ◽  
Parasitic Worm ◽  
Mouse Mast Cell ◽  
Protein Kinase C Isoforms

scholarly journals Role of protein subunits in Proteus rettgeri penicillin G acylase.

1985 ◽  
Vol 163 (3) ◽  
pp. 1279-1281 ◽  
Author(s):  
G O Daumy ◽  
D Danley ◽  
A S McColl
Keyword(s):  
Penicillin G Acylase ◽  
Penicillin G ◽  
Protein Subunits

Altered heart rate control in transgenic mice carrying the KCNJ6 gene of the human chromosome 21

2008 ◽  
Vol 33 (2) ◽  
pp. 230-239 ◽  
Author(s):  
Jacques M. Lignon ◽  
Zoë Bichler ◽  
Bruno Hivert ◽  
François E. Gannier ◽  
Pierre Cosnay ◽  
...  
Keyword(s):  
Human Chromosome ◽  
Heart Defects ◽  
Chromosome 21 ◽  
P Wave ◽  
Altered Expression ◽  
Protein Subunits ◽  
Human Chromosome 21 ◽  
Pr Interval ◽  
Cardiac Regulation

Congenital heart defects (CHD) are common in Down syndrome (DS, trisomy 21). Recently, cardiac sympathetic-parasympathetic imbalance has also been documented in DS adults free of any CHD. The KCNJ6 gene located on human chromosome 21 encodes for the Kir3.2/GIRK2 protein subunits of G protein-regulated K+ (KG) channels and could contribute to this altered cardiac regulation. To elucidate the role of its overexpression, we used homozygous transgenic (Tg+/+) mice carrying copies of human KCNJ6. These mice showed human Kir3.2 mRNA expression in the heart and a 2.5-fold increased translation in the atria. Phenotypic alterations were assessed by recording electrocardiogram of urethane anesthetized mice. Chronotropic responses to direct (carbachol) and indirect (methoxamine) muscarinic stimulation were enhanced in Tg+/+ mice with respect to wild-type (WT) mice. Alternating periods of slow and fast rhythm induced by CCPA (2-chloro- N-cyclopentyl-adenosine) were amplified in Tg+/+ mice, resulting in a reduced negative chronotropic effect. These drugs reduced the atrial P wave amplitude and area. P wave variations induced by methoxamine and CCPA were respectively increased and reduced in the Tg+/+ mice, while PR interval and ventricular wave showed no difference between Tg+/+ and WT. These results indicate that Tg+/+ mice incorporating the human KCNJ6 exhibit altered Kir3.2 expression and responses to drugs that would activate KG channels. Moreover, these altered expression and responses are limited to sino-atrial node and atria that normally express large amounts of KG channels. These data suggest that KCNJ6 could play an important role in altered cardiac regulation in DS patients.

scholarly journals Repositioning the Sm-binding site in S. cerevisiae telomerase RNA reveals RNP organizational flexibility and Sm-directed 3′-end formation

2017 ◽  
Author(s):  
Evan P. Hass ◽  
David C. Zappulla
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Binding Site ◽  
Strong Evidence ◽  
Binding Sites ◽  
Telomerase Rna ◽  
Telomeric Dna ◽  
Protein Subunits ◽  
Yeast Telomerase ◽  
Rna Biogenesis

ABSTRACTTelomerase RNA contains a template for synthesizing telomeric DNA by reverse transcription and has been proposed to act as a flexible scaffold for holoenzyme protein subunits in the RNP. In Saccharomyces cerevisiae, the telomerase subunits Est1 and Ku bind to the telomerase RNA, TLC1, and it has been shown that these proteins still function when their binding sites are repositioned within the RNA. TLC1 is also bound by the Sm7 protein complex, which is required for stabilization of the predominant, non-polyadenylated (poly(A)–) TLC1 isoform. Here, we first show that Sm7 can perform this function even when its binding site is repositioned via circular permutation to several different positions within TLC1, further supporting the conclusion that the telomerase holoenzyme is organizationally flexible. Next, we tested the hypothesis that the location of the Sm7-binding site relative to the 3′ end is contrastingly important. When we moved the Sm site to locations 5′ of its native position, we observed that this stabilized shorter forms of poly(A)– TLC1 in a manner precisely corresponding to how far upstream the Sm site was moved. This provides strong evidence that the location of Sm7 binding to TLC1 controls where the mature poly(A)– 3′ end is formed. In summary, our results show that Sm7 and the 3′ end of yeast telomerase RNA comprise an organizationally flexible module within the telomerase RNP and provide insights into the mechanistic role of Sm7 in telomerase RNA biogenesis.

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