The Heptameric SmAP1 and SmAP2 Proteins of the Crenarchaeon Sulfolobus Solfataricus Bind to  Common and Distinct RNA Targets

Märtens, Birgit; Bezerra, G.A.; Kreuter, Mathias Josef; Grishkovskaya, Irina; Manica, Andrea; Arkhipova, Valentina; Djinović-Carugo, Kristina; Bläsi, Udo

doi:10.3390/life5021264

Cited by 18 publications

(44 citation statements)

References 50 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The genes encoding the SmAP1 and SmAP2 proteins were abutted at the 3΄end with a 10×His-tag encoding sequence. Transcription of the genes from the virus-derived pMJ05-vector was controlled by an arabinose-inducible promoter in Sso PH1-16 ( 26 ). When compared with endogenous protein levels, this expression system permits a 3- to 10-fold overproduction of the respective proteins ( 48 , 55 ).…”

Section: Resultsmentioning

confidence: 99%

“…The Sso strains PH1-16(pMJ05-SmAP1-His), PH1-16(pMJ05-SmAP2-His) ( 26 ) and PH1-16(pMJ05-DnaG-His) were generated as described ( 48 ) ( Supplementary Data ). For expression of the plasmid borne genes, the strains PH1-16(pMJ05), harboring the backbone vector (mock control), PH1-16(pMJ05-SmAP1-His), PH1-16(pMJ05-SmAP2-His) and PH1-16(pMJ05-DnaG-His) were grown at 75°C in arabinose-containing Brock's medium.…”

Section: Methodsmentioning

confidence: 99%

“…The SmAP1 coding gene was cloned as described in Supplementary Data . DnaG-His, SmAP1-His and SmAP2-His were purified as previously described ( 26 , 49 ). For Co-IP with purified components, 50 pmol of DnaG-His was either incubated alone (mock control) or SmAP1-His and SmAP2-His were incubated together with 50 pmol DnaG-His, respectively, in 200 μl Co-IP buffer (50 mM Tris–HCl pH 6.0, 100 mM KCl; 5% glycerol, 1 mM MgCl 2 and 0.1% Triton X-100) for 10 min at 65°C.…”

Section: Methodsmentioning

confidence: 99%

“…While bacterial and eukaryotic Lsm family members have been studied in more detail ( 2 , 4 , 5 , 14 ), the function of Sm-like archaeal proteins (SmAPs) remains poorly understood. Like other Sm/Lsm proteins, the SmAPs are composed of an N-terminal α-helix and five β-strands forming a continuous Sm-fold ( 22 – 26 ). However, the homo-heptameric SmAPs display characteristic differences from bacterial and eukaryotic Lsm proteins, e.g.…”

Section: Introductionmentioning

confidence: 99%

“…SmAP1 and SmAP2 show 50% similarity, whereas they share only 30% similarity with SmAP3. In Sso different classes of non-coding RNAs and mRNAs were identified that interact either with SmAP1 or SmAP2 or with both proteins ( 26 ). The large number of associated intron-containing tRNAs and rRNA modifying RNAs suggested as well a role of these SmAPs in tRNA/rRNA processing ( 26 ).…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

The SmAP1/2 proteins of the crenarchaeon Sulfolobus solfataricus interact with the exosome and stimulate A-rich tailing of transcripts

Märtens

Hou

Amman

et al. 2017

Nucleic Acids Research

Self Cite

View full text Add to dashboard Cite

The conserved Sm and Sm-like proteins are involved in different aspects of RNA metabolism. Here, we explored the interactome of SmAP1 and SmAP2 of the crenarchaeon Sulfolobus solfataricus (Sso) to shed light on their physiological function(s). Both, SmAP1 and SmAP2 co-purified with several proteins involved in RNA-processing/modification, translation and protein turnover as well as with components of the exosome involved in 3΄ to 5΄ degradation of RNA. In follow-up studies a direct interaction with the poly(A) binding and accessory exosomal subunit DnaG was demonstrated. Moreover, elevated levels of both SmAPs resulted in increased abundance of the soluble exosome fraction, suggesting that they affect the subcellular localization of the exosome in the cell. The increased solubility of the exosome was accompanied by augmented levels of RNAs with A-rich tails that were further characterized using RNASeq. Hence, the observation that the Sso SmAPs impact on the activity of the exosome revealed a hitherto unrecognized function of SmAPs in archaea.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

The SmAP1/2 proteins of the crenarchaeon Sulfolobus solfataricus interact with the exosome and stimulate A-rich tailing of transcripts

Märtens

Hou

Amman

et al. 2017

Nucleic Acids Research

Self Cite

View full text Add to dashboard Cite

show abstract

Modification of translation factor aIF5A from Sulfolobus solfataricus

et al. 2018

View full text Add to dashboard Cite

Eukaryotic eIF5A and its bacterial orthologue EF-P are translation elongation factors whose task is to rescue ribosomes from stalling during the synthesis of proteins bearing particular sequences such as polyproline stretches. Both proteins are characterized by unique post-translational modifications, hypusination and lysinylation, respectively, which are essential for their function. An orthologue is present in all Archaea but its function is poorly understood. Here, we show that aIF5A of the crenarchaeum Sulfolobus solfataricus is hypusinated and forms a stable complex with deoxyhypusine synthase, the first enzyme of the hypusination pathway. The recombinant enzyme is able to modify its substrate in vitro resulting in deoxyhypusinated aIF5A. Moreover, with the aim to identify the enzyme involved in the second modification step, i.e. hypusination, a set of proteins interacting with aIF5A was identified.Electronic supplementary materialThe online version of this article (10.1007/s00792-018-1037-4) contains supplementary material, which is available to authorized users.

show abstract

An Experimental Tool to Estimate the Probability of a Nucleotide Presence in the Crystal Structures of the Nucleotide–Protein Complexes

et al. 2017

View full text Add to dashboard Cite

A correlation between the ligand-protein affinity and the identification of the ligand in the experimental electron density maps obtained by X-ray crystallography has been tested for a number of RNA-binding proteins. Bacterial translation regulators ProQ, TRAP, Rop, and Hfq together with their archaeal homologues SmAP have been used. The equilibrium dissociation constants for the N-methyl-anthraniloyl-labelled adenosine and guanosine monophosphates titrated by the proteins have been determined by the fluorescent anisotropy measurements. The estimated stability of the nucleotide-protein complexes has been matched with a presence of the nucleotides in the structures of the proposed nucleotide-protein complexes. It has been shown that the ribonucleotides can be definitely identified in the experimental electron density maps at equilibrium dissociation constant <10 μM. At K of 20-40 μM, long incubation of the protein crystals in the nucleotide solution is required to obtain the structures of the complexes. The complexes with K value higher than 50 μM are not stable enough to survive in crystallization conditions.

show abstract

The Heptameric SmAP1 and SmAP2 Proteins of the Crenarchaeon Sulfolobus Solfataricus Bind to Common and Distinct RNA Targets

Cited by 18 publications

References 50 publications

The SmAP1/2 proteins of the crenarchaeon Sulfolobus solfataricus interact with the exosome and stimulate A-rich tailing of transcripts

The SmAP1/2 proteins of the crenarchaeon Sulfolobus solfataricus interact with the exosome and stimulate A-rich tailing of transcripts

Modification of translation factor aIF5A from Sulfolobus solfataricus

An Experimental Tool to Estimate the Probability of a Nucleotide Presence in the Crystal Structures of the Nucleotide–Protein Complexes

Contact Info

Product

Resources

About