Display Settings:

Items per page

Results: 6

1.
FIG. 4.

FIG. 4. From: Bacillus subtilis YhaM, a Member of a New Family of 3?-to-5? Exonucleases in Gram-Positive Bacteria.

Growth curves of RNase mutants. The pnpA rnr strain (solid circles) and the pnpA rnr yhaM strain (open circles) were grown at 37°C in LB medium. Values are averages from two experiments.

Irina A. Oussenko, et al. J Bacteriol. 2002 November;184(22):6250-6259.
2.
FIG. 3.

FIG. 3. From: Bacillus subtilis YhaM, a Member of a New Family of 3?-to-5? Exonucleases in Gram-Positive Bacteria.

(A) Divalent-cation dependence of YhaM activity. The uniformly labeled 110-nt RNA substrate (A) or the 5′-end-labeled 38-nt DNA substrate (B) was incubated with purified YhaM in the presence of either the indicated divalent cation at 1 mM or no divalent cation (−). Lanes C, control lanes containing the substrate incubated without YhaM and without divalent cations. Migration of mononucleotides is indicated.

Irina A. Oussenko, et al. J Bacteriol. 2002 November;184(22):6250-6259.
3.
FIG. 2.

FIG. 2. From: Bacillus subtilis YhaM, a Member of a New Family of 3?-to-5? Exonucleases in Gram-Positive Bacteria.

Directionality of exoribonuclease activity. (A) Predicted secondary structure within the 187-nt RNA substrate. The 3′ end of the ∼110-nt RNA is located in the loop region, as indicated. (B) YhaM-mediated degradation of 187- and 110-nt RNAs. Migration of the full-length substrates is indicated on the right. The arrow at right indicates a decay intermediate resulting from a block to 3′-to-5′ exonuclease activity at the base of the stem structure. Lanes C, control lanes containing an untreated RNA substrate.

Irina A. Oussenko, et al. J Bacteriol. 2002 November;184(22):6250-6259.
4.
FIG. 5.

FIG. 5. From: Bacillus subtilis YhaM, a Member of a New Family of 3?-to-5? Exonucleases in Gram-Positive Bacteria.

Comparison of YhaM and CBF1. (A) SDS-10% PAGE gel of His-tagged YhaM and CBF1 proteins isolated from an Ni2+-NTA column (Ni) or blotted onto a PVDF membrane and eluted under nonreducing (NR) or reducing (R) conditions. In the case of CBF1, the upper and lower bands (indicated by asterisks to the left of the bands in lane 4) were isolated from the PVDF membrane separately. Molecular sizes (in kilodaltons) of protein markers (lane M) are given on the left. (B) RNase activities of YhaM and CBF1 proteins, showing Mn2+ dependence and increased activity of CBF1 when isolated under reducing conditions. Arrow at left indicates migration of mononucleotide product.

Irina A. Oussenko, et al. J Bacteriol. 2002 November;184(22):6250-6259.
5.
FIG. 1.

FIG. 1. From: Bacillus subtilis YhaM, a Member of a New Family of 3?-to-5? Exonucleases in Gram-Positive Bacteria.

Purification of YhaM. (A) Silver-stained SDS-10% PAGE gel with a purified fraction from the Mono Q column (lane 1) and an isolated 39-kDa protein with exoribonuclease activity (lane 2). Arrow indicates the migration of the YhaM protein. Molecular sizes (in kilodaltons) of protein markers (lane M) are given on the left. (B) Exoribonuclease activity of protein bound to Ni2+-NTA resin, from E. coli strains carrying a His tag expression vector with the yhaM coding sequence (filled circles) or the vector alone (open circles). Samples were taken at the time of isopropyl-β-d-thiogalactopyranoside (IPTG) addition (0 h) and 2 h thereafter (2 h). Activity was assayed in the presence of Mn2+ or Mg2+. Lane C, control lane containing an untreated RNA substrate. Arrow at right indicates labeled mononucleotide product. Upon overexposure, a band at this position is also visible in the lane next to the control lane. The gel was a 20% denaturing polyacrylamide gel.

Irina A. Oussenko, et al. J Bacteriol. 2002 November;184(22):6250-6259.
6.
FIG. 6.

FIG. 6. From: Bacillus subtilis YhaM, a Member of a New Family of 3?-to-5? Exonucleases in Gram-Positive Bacteria.

Comparison of the crystal structure of the RPA32 subunit of replication protein A (A) (11) and the YhaM OB-fold domain model (B). Residues predicted to stack with single-stranded DNA bases in RPA32 (11) and their equivalent residues in YhaM are numbered and shown as ball-and-stick models. N and C termini are labeled. (C) Multiple alignment of 11 YhaM homologues. The OB-fold domain is shown in orange, with YhaM W51 and Y76 positions marked in yellow. The HD domain is shown in green, with a possible extension in light green. The last line indicates sequence conservation, with completely conserved positions marked by asterisks and partially conserved positions marked by periods or colons, indicating lesser or greater conservation, respectively. The alignment was calculated with CLUSTAL W (47) using sequences that contained both OB-fold and HD domains. Sequences are identified by their National Center for Biotechnology Information gene identifier numbers and organism abbreviations as follows: Bsu, B. subtilis; Sau, S. aureus; Lin, Listeria innocua; Lmo, Listeria monocytogenes; Spn, Streptococcus pneumoniae; Spy, Streptococcus pyogenes; Bha, Bacillus halodurans; Lla, Lactococcus lactis; Cac, C. acetobutylicum; Fis, F. islandicum.

Irina A. Oussenko, et al. J Bacteriol. 2002 November;184(22):6250-6259.

Display Settings:

Items per page

Supplemental Content

Recent activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...
Write to the Help Desk