Coleman AW, Preparata RM, Mehrotra B, et al.

Derivation of the secondary structure of the ITS-1 transcript in volvocales and its taxonomic correlations
PROTIST 149 (2): 135-146 MAY 1998

Abstract:
Knowledge of secondary structure, formed by the gene spacer regions of the primary transcript of nuclear rDNA cistrons, is lacking for most phyla of eukaryotes. We have sequenced the first internal transcribed spacer region (ITS-1) of multiple representatives of the Volvocales, and from comparisons of these, derived a secondary structure common to the entire group. The secondary structure model is supported by numerous compensating base pair changes located within the paired regions of the stem-loops. Within the morphological species, such as those of Astrephomene and Gonium, the three basal nucleotide pairs of helices are highly conserved in primary sequence, and the single stranded region rich in CCAA is identical in sequence, even when isolates come from all continents of the earth. In other Volvocacean species known to include many pairs of mating types, this same level of conservation is found to correlate with the mating subgroups of the species. Thus a comparable degree of sequence similarity appears to characterize all isolates of a "biological" species; this is valid for taxonomic species only where the biological and taxonomic species levels coincide. In addition, the ITS-1 contains information useful for population analyses, and spacer secondary structure may have additional phylogenetic utility at the level of class or subclass when that information becomes available for other protistan groups.

Nozaki H, Ohta N, Yamada T, et al.

Characterization of rbcL group IA introns from two colonial volvocalean species (Chlorophyceae)
PLANT MOL BIOL 37 (1): 77-85 MAY 1998

Abstract:
Group I introns were reported for the first time in the large subunit of Rubisco (rbcL) genes, using two colonial green algae, Pleodorina californica and Gonium multicoccum (Volvocales). The rbcL gene of P. californica contained an intron (PlC intron) of 1320 bp harboring an open reading frame (ORF). The G. multicoccum rbcL gene had two ORF-lacking introns of 549 (GM1 intron) and 295 (GM2 intron) base pairs. Based on the conserved nucleotide sequences of the secondary structure, the PlC and GM1 introns were assigned to group IA2 whereas the GM2 intron belonged to group IA1. Southern hybridization analyses of nuclear and chloroplast DNAs indicated that such intron-containing rbcL genes are located in the chloroplast genome. Sequencing RNAs from the two algae revealed that these introns are spliced out during mRNA maturation. In addition, the PlC and GM1 introns were inserted in the same position of the rbcL exons, and phylogenetic analysis of group IA introns indicated a close phylogenetic relationship between the PlC and GM1 introns within the lineage of bacteriophage group IA2 introns. However, P. californica and G. multicoccum occupy distinct clades in the phylogenetic trees of the colonial Volvocales, and the majority of other colonial volvocalean species do not have such introns in the rbcL genes. Therefore, these introns might have been recently inserted in the rbcL genes independently by horizontal transmission by viruses or bacteriophage.

Haller K, Fabry S

Brefeldin A affects synthesis and integrity of a eukaryotic flagellum
BIOCHEM BIOPH RES CO 242 (3): 597-601 JAN 26 1998

Abstract:
Eukaryotic flagella and cilia are highly dynamic organelles. In green algae like Chlamydomonas reinhardtii, flagella absorption and resynthesis is a normal process during the vegetative cell cycle. Rapid regeneration also occurs after stress-induced shedding of flagella. Ca2+ ions, protein synthesis, and a kinase activity are the main factors known to affect resynthesis. Recently, we have detected that certain small G proteins (Ypt/Rab) and a GTPase regulator (GDP dissociation inhibitor), known as regulatory elements of intracellular vesicle transport, are present in flagellar membranes of green algae, raising the possibility that the organelle's synthesis and/or integrity depends on functional membrane traffic. In this study, we examined the effect of brefeldin A (BFA), an inhibitor of intracellular membrane flow and Golgi function in animal and plant cells, on flagella regeneration in the colonial green alga Gonium pectorale. We show that low BFA concentrations (<1 mu g/ml) inhibit flagella outgrowth, while higher amounts cause dose-dependent deflagellation and cell death. Our findings provide experimental evidence for a direct connection between intracellular transport and eukaryotic flagella synthesis. (C) 1998 Academic Press.