Heterogeneity of the ribosomal protein pattern in mycelium of Streptomyces species

Abstract

The pattern of ribosomal proteins of 7 Streptomyces species was studied. Proteins were separated by polyacrylamide gel electrophoresis (PAGE) using two different electrophoretic systems, and also by reverse‐phase high‐performance liquid chromatography (HPLC). By these 3 methods, clear differences were observed, in several proteins (absent in some species and present in others), with some species showing more closely related ribosomal protein patterns than others. It is suggested that the application of the present methods may be of some potential in a taxonomic context.

Cloning, sequencing and transcriptional analysis of a Streptomyces coelicolor operon containing the rplM and rpsI genes encoding ribosomal proteins ScoL13 and ScoS9

“…Ribosomes from S. coelicolor were isolated (Quiro s et al 1989) and total 70S ribosomal proteins were puri®ed (Fierro et al 1987) and separated by two-dimensional PAGE, using a nonequilibrium pH gel electrophoresis system (NEPHGE) for the ®rst dimension (VanBogelen et al 1990;Blanco et al 1994). Proteins from several spots were digested with lysyl-endopeptidase, and the resulting peptides were separated by high-performance liquid chromatography (HPLC) as described by Kawasaki et al (1990).…”

“…Important aspects of primary metabolism, such as the machinery involved in protein synthesis, have not been studied in such great detail. Dierences in the ribosomal protein pattern among several streptomycete species have been described (Fierro et al 1987;Ochi 1992) and changes in the pattern of ribosomal proteins during spore germination (Mikulik et al 1984;Quiro s et al 1989) and during the transition from substrate to aerial mycelium (Quiro s et al 1992;Blanco et al 1994) have also been noted. Only a few genes encoding Streptomyces ribosomal proteins have been cloned and sequenced and they correspond to only a few operons: the L11 and L10 operons (Blanco et al 1992(Blanco et al , 1994Parra et al 1992;Ruengjitchawalya et al 1993;Katayama et al 1996), the L34 operon (Calcutt and Schmidt 1992), the spc operon (Hale et al 1995) and the alpha operon (Cho et al 1996).…”

Synthesis of ribosomal proteins during growth of Streptomyces coelicolor

“…Ribosomes were prepared from S. coelicotor mycelia grown in MG medium as described before. Mycelia were broken with ultrasound (10 pulses of 30 s each with intermittent cooling) in a 150W ultrasonic disintegrator, Ribosomes were purified by successive ulracentrifugation steps through high-salt buffer and sucrose as described (Quiros et ai, 1989), Ribosomat proteins were extracted from purified ribosomes with glacial acetic acid (Fierro et al, 1987).…”

Structural and Functional Analysis of Ribosomal Subunits from Vegetative Mycelium and Spores of Streptomyces antibioticus

“…Proteins were extracted from ribosomes as described previously (Fierro et al, 1987). Samples of ribosomal subunit proteins from dormant spores or vegetative mycelium, containing 100 pg protein in buffer D, were fractionated by reverse-phase high-performance liquid chromatography (RP-HPLC) using the method of Kerlavage et al (1983) with some minor modifications (Fierro el al., 1987).…”

“…SDS-PAGE was done using 8-20% (w/v, acrylamide) gradient gel. The method used was based on that described by Laemmli (1970), with modifications (Fierro et al, 1987). The following marker proteins (Pharmacia low molecular mass kit) were used : phosphorylase b (molecular mass 94000 kDa), bovine serum albumin (67000 kDa), ovalbumin (43 000 kDa), carbonic anhydrase (30000 kDa), soybean trypsin inhibitor (20 100 kDa) and P-lactalbumin (14400 kDa).…”