Molecular evolution of the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO)

Abstract

The evolutionary relationship of the RuBisCO large subunit gene(s) (rbcL) of several prokaryotes was examined using the technique of heterologous DNA hybridization. Restriction fragments of cloned rbcL from Anacystis nidulans 6301, Chlamydornonas reinhardtii, Rhodospirillum rubrum, and maize were nick-translated and used as probes. The C. reinhardtii and maize probes hybridized with restriction fragment(s) only from cyanobacteria: Agmenellum quadruplicatum, Fremyella diplosiphon, and Mastigocladus laminosus. In addition, the A. nidulans probe hybridized with restriction fragment (s) from Alcaligenes eutrophus, Chromatium vinosum, Nitrobacter hamburgensis, Paracoccus denitrificans, Pseudomonas oxalaticus, Rhodomicrobium oannielii, Rhodopseudomonas capsulata, Rhodopseudornonas palustris, Rhodopseudomonas sphaeroides, Thiobacillus intermedius, Thiobacillus neapolitanus, and Thiothrix nivea. The elucidated fragment of Rhodopseudomonas species is presumably for the Form I RuBisCO LSU of these organisms. The R. rubrum probe hybridized only to a restriction fragment(s) from R. capsulata, R. palustris, R. sphaeroides, T. neapolitanus, and T. nivea. The fragment(s) of Rhodopseudornonas species is the Form II rbcL of these organisms.'Fhe restriction fragments of T. neapolitanus and T. nivea were also different from those elucidated by the A. nidulans probe, suggesting the presence of a second (different) rbcL in these organisms. Positive hybridization was not obtained using any of the probes with DNA from Beggiatoa alba, Chlorobium vibrioforme or Chloroflexus aurantiacus. It appears that all rbcL have evolved from a common ancestor. Our data are consistent with and supportive of the evolutionary scheme for RuBisCO proposed by Akazawa, Takabe, and Kobayashi [1].

An immunoprobe to measure Rubisco concentrations and maximal photosynthetic rates of individual phytoplankton cells

“…One reason that it was possible to select for a species-independent Rubisco probe is that the large subunit protein is highly conserved in an evolutionary context (Miziorko and Lorimer 19 8 3). Immunological cross-reactivity (Table l), hybrid reconstitution (Andrews and Lorimer 1987), heterologous hybridization (Shively et al 1986), and amino acid sequences (Miziorko and Lorimer 1983) all suggest that the large subunit of higher plants, green algae, and cyanobacteria is conservative, particularly at the active sites. Taxa-specific differences have been reported for other anti-Rubisco antisera (Plumley et al 1986;Newman and Cattolico 1987b); it should be noted that these polyclonal antisera were not affinity purified to conservative sites.…”

Cloning and expression of the ?-ribulose-1,5-bisphosphate carboxylase/oxygenase form II gene from Thiobacillus intermedius in Escherichia coli

“…intermedius cells were harvested, resuspended, washed and lysed as previously described except that Triton X-100 was replaced with Brij 58 and sodium deoxycholate at final concentrations of 2.0% (w/v) and 0.2% (w/v), respectively [21]. Plasmid and genomic DNA were purified by cesium chloride-ethidium bromide density gradients [21]. DNA digestion and Southern blotting were accomplished as previously described [21].…”

“…Plasmid and genomic DNA were purified by cesium chloride-ethidium bromide density gradients [21]. DNA digestion and Southern blotting were accomplished as previously described [21].…”

“…Gene probes for phosphoribulokinase, fructose bisphosphatase and glyceraldehyde phosphate dehydrogenase were also tested ( Table 1). Probe preparation and hybridization were carried out as previously described [21]. Several positive plaques were obtained when the genomic library was screened with pBSSP720.…”

Insertion Mutation of the Form I cbbL Gene Encoding Ribulose Bisphosphate Carboxylase/Oxygenase (RuBisCO) in Thiobacillus neapolitanus Results in Expression of Form II RuBisCO, Loss of Carboxysomes, and an Increased CO 2 Requirement for Growth

“…denitrificans expresses both a form I and a form II RuBisCO when grown anaerobically with nitrate as the electron acceptor (7). The genes for both form I and form II (cbbM) RuBisCO have been demonstrated in several other thiobacilli, including T. neapolitanus, T. intermedius, T. ferrooxidans, and T. thiooxidans, via heterologous hybridization using specific DNA probes (28). The cbbM gene from one of these thiobacilli, T. intermedius, has been isolated and expressed in Escherichia coli (32).…”