Name: Proteins identified by LC-MS/MS analysis to associate with RecJ4-StrepII (vs. empty vector).
Keywords: Recj4

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Proteins identified by LC-MS/MS analysis to associate with RecJ4-StrepII (vs. empty vector).

ABSTRACT: RecJ4 association with RecJ3, RNase J and Cdc48A investigated by pullingown strep tagged-RecJ4 with its partner proteins

INSTRUMENT(S): LTQ Orbitrap XL

ORGANISM(S): Haloferax Volcanii Ds2

SUBMITTER: Swathi Dantuluri

LAB HEAD: Julie Maupin-Furlow

PROVIDER: PXD019933 | Pride | 2023-06-22

REPOSITORIES: Pride

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RNase J, RecJ3, RNase J, RecJ3, Cdc48a and Cas7 co-purify with RecJ4-StrepII in the presence of γATP. and Cas7 co-purify with RecJ4-StrepII in the presence of γATP.

Project description:detecting core complex formation for RecJ3/4 and RNase J form a core complex.

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Proteins associated with Cdc48A identified by LC-MS\MS data

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Project description:H. volcanii Cdc48a, RecJ3/4 and RNase J associate with Ubl-coated beads. A) Strategy to isolate H. volcanii proteins that bind agarose beads charged with monomeric Ubl (vs. BSA) as bait. Cell lysate was from triplicate cultures of H. volcanii NH02-pJAM957. B) Left, Non-reducing SDS-PAGE of H. volcanii proteins that bound BSA- (control, lane 1) vs. Ubl- (lane 2) decorated beads in the presence of ATP. Gels were stained with SYPRO Ruby. Red bar, HMW(250kDa) and LMW(50kDa) (high and low molecular weight) regions of gel excised from lanes 1 and 2 for LC- MS/MS analysis. Right, Proteins identified at > 99.9% probability and < 0.1 % false discovery rate (FDR) in the Ubl (not BSA) samples. Similar trend was observed in independent experiment. ATP was required for this association. Theor. Mr, theoretical molecular mass based on genome sequence. Normalized total spectra of sample minus control is indicated.

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Project description:The data comes from a study, where three-spined sticklebacks (Gasterosteus aculeatus) were exposed to di-n-butyl phthalate (DBP) and 17α ethinyl-oestradiol (EE2) at nominal concentrations 35 μg/L and 40 ng/L, respectively, for four days. The aim of the study was to obtain insight into the acute transcriptional responses putatively associated with endocrine disruption. RNA samples from the testes of eight individuals fish per treatment (including solvent controls, exposed only to DMSO) were used in the microarray analysis, covering the expression of approximately 21000 genes.

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