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Characterization of molecular partners and physiological functions of the conserved RNA binding protein Rbp3

ABSTRACT: The RNA recognition motif (RRM) domain proteins are crucial RB (Rbps) in all domains of life. In cyanobacteria, proteins with a single RRM domain were suggested to be involved in mRNA targeting to the thylakoid membrane and the acclimation to certain stress conditions. The model cyanobacterium Synechocystis sp. PCC 6803 contains a small family of 3 genes encoding RRM-domain containing Rbps, Rbp1, Rbp2 and Rbp3. With a focus on Rbp3, here, we addressed the molecular details of its functions and the associated physiological consequences. To elucidate the set of interacting proteins, co-immunoprecipitation experiments were performed with a strain expressing an Rbp3 with a triple FLAG tag added to its C-terminus.

INSTRUMENT(S): LTQ Orbitrap Elite

ORGANISM(S): Synechocystis Sp. Pcc 6803 (ncbitaxon:1148)

SUBMITTER: Wolfgang R. Hess

PROVIDER: MSV000092937 | MassIVE |

SECONDARY ACCESSION(S): PXD045584

REPOSITORIES: MassIVE

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The RRM domain-containing protein Rbp3 interacts with ribosomes and the 3’ ends of mRNAs encoding photosynthesis proteins

Project description:Gene expression changes were monitored in cultures of the cyanobacterium Synechocystis sp. PCC 6803 at 30 min after a shift from cultivation under standard light conditions (50 µmol photons m-2 s-1) to high light (HL; 250 µmol photons m-2 s-1) and 1 h after a shift from the standard growth temperature of 30°C to 20°C (cold). Acclimation to shifts in light and temperature is critical for photoautotrophic organisms such as cyanobacteria. The RRM domain proteins Rbp2 and Rpb3 are RNA-binding proteins that bind specific mRNAs and are involved in the localization of these mRNAs to the thylakoid membrane. Here, we investigated the effects of Rpb2 and Rbp3 on gene expression changes during the acclimation responses to an increase in light intensity and a decrease in temperature. We analyzed cultures of the wild-type (WT) and of the Δrbp2Δrbp3 double mutant, which lacks Rbp2 and Rbp3. We found that the transcript levels of psaA and psaB genes were significantly decreased in cells of the Δrbp2Δrbp3 double mutant upon cold stress and 30 min of high light compared to wild type. In addition, the transcript levels of Rbp1, another RRM domain-containing protein, were greatly increased.

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The proteome of Synechocystis sp. PCC 6803 cells grown under light-activated heterotrophic growth

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2015-03-03 | PXD000980 | Pride

In vivo recruitment analysis and a mutant strain without any group 2 σ factors reveal roles of different σ factors in cyanobacteria

Project description:In eubacteria, replacement of one σ factor in the RNA polymerase (RNAP) holoenzyme by another one changes the transcription pattern. Cyanobacteria are eubacteria characterized by oxygenic photosynthesis and they typically encode numerous group 2 σ factors that closely resemble the essential primary σ factor. A mutant strain of the model cyanobacterium Synechocystis sp. PCC 6803 without functional group 2 σ factors (named as ΔsigBCDE) could not acclimate to heat, high salt, or bright light stress but in standard conditions ΔsigBCDE grew only 9% slower than the control strain. One-fifth of the genes in ΔsigBCDE were differently expressed compared to the control strain in standard growth conditions and several physiological changes in photosynthesis, and pigment and lipid compositions were detected. To directly analyze the σ factor content of RNAP holoenzyme in vivo, a His-tag was added to the γ subunit of RNAP in Synechocystis and RNAPs were collected. The results revealed that all group 2 σ factors were recruited by RNAP in standard conditions, but recruitment of SigB and SigC increased in heat stress, SigD in bright light, SigE in darkness and SigB, SigC and SigE in high salt, explaining the poor acclimation of ΔsigBCDE to these stress conditions. Cells from cyanobacteria Synechocystis sp. PCC 6803 named as control strain (CS) and a mutant strain without any functional group 2 sigma factors, ΔsigBCDE, were harvested (A730=1, 40 mL) directly from standard growth conditions (continuous illumination at the PPFD of 40 µmol m-2s-1, 32°C, ambient CO2). From three to four independent experiments were performed at each conditions.

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2021-11-10 | PXD023181 | Pride

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