Nozaki H, Itoh M, Sano R, et al.

Phylogenetic relationships within the colonial volvocales (Chlorophyta) inferred from rbcL gene sequence data
J PHYCOL 31 (6): 970-979 DEC 1995

Abstract:
The chloroplast-encoded large subunit of the ribulose-1, 5-bisphosphate carboxylase/oxygenase (rbcL) gene was sequenced from 20 species of the colonial Volvocales (the Volvacaceae, Goniaceae, and Tetrabaenaceae) in order to elucidate phylogenetic relationships within the colonial Volvocales. Eleven hundred twenty-eight base pairs In the coding regions of the (rbcL) gene were analyzed by the neighbor-joining (NJ) method using three kinds of distance estimations, as well as by the maximum parsimony (MP) method. A large group comprising all the anisogamous and oogamous volvocacean species was resolved in the MP tree as well as in the NJ trees based on overall and synonymous substitutions. In all the trees constructed, Basichlamys and Tetrabaena (Tetrabaenaceae) constituted a very robust phylogenetic group. Although not supported by high bootstrap values, the MP tree and the NJ tree based on nonsynonymous substitutions indicated that the Tetrabaenaceae is the sister group to the large group comprising the Volvocaceae and the Goniaceae. In addition, the present analysis strongly suggested that Pandorina and Astrephomene are monophyletic genera whereas Eudorina is nonmonophyletic. These results are essentially consistent with the results of the recent cladistic analyses of morphological data. However, the monophyly of the Volvocaceae previously supported by four morphological synapomorphies is found only in the NJ tree based on nonsynonymous substitutions (with very low bootstrap values). The genus Volvox was clearly resolved as a polyphyletic group with V. rousseletii Pocock separated from other species of volvox in the rbcL gene comparisons, although this genus represents a monophyletic group in the previous morphological analyses. Furthermore, none of the rbcL gene trees supported the monophyly of the Goniaceae; Astrephomene was placed in various phylogenetic positions.

 

Memon A, Hwang SB, Deshpande N, et al.

Novel aspects of the regulation of a cDNA (Arf1) from Chlamydomonas with high sequence identity to animal ADP-ribosylation factor 1
PLANT MOL BIOL 29 (3): 567-577 NOV 1995

Abstract:
ADP-ribosylation factor (ARF) is a highly conserved, low molecular mass (ca. 21 kDa) GTP-binding protein that has been implicated in vesicle trafficking and signal transduction in yeast and mammalian cells. However, little is known of ARF in plant systems. A putative ARF polypeptide was identifed in subcellular fractions of the green alga Chlamydomonas reinhardtii, based on [P-32]GTP binding and immunoblot assays. A cDNA clone was isolated from Chlamydomonas (Arfl), which encodes a 20.7 kDa protein with 90% identity to human ARF1. Northern blot analyses showed that levels of Arfl mRNA are highly regulated during 12 h/12 h light/dark (LD) cycles. A biphasic pattern of expression was observed: a transient peak of Arfl mRNA occurred at the onset of the light period, which was followed ca. 12 h later by a more prominent peak in the early to mid-dark period. When LD-synchronized cells were shifted to continuous darkness, the dark-specific peak of Arfl mRNA persisted, indicative of a circadian rhythm. The increase in Arfl mRNA at the,beginning of the light period, however, was shown to be light-dependent, and, moreover, dependent on photosynthesis, since it was prevented by DCMU. We conclude that the biphasic pattern of Arfl mRNA accumulation during LD cycles is due to regulation by two different factors, light (which requires photosynthesis) and the circadian clock. Thus, these studies identify a novel pattern of expression for a GTP-binding protein gene.

 

Imaizumi M, Doida Y

Cyclic AMP is a signal for repression of differentiation into gametes in Micrasterias thomasiana var notata
PLANT SCI 112 (1): 33-42 NOV 24 1995

Abstract:
This study investigates the effects of adenosine 3':5'-cyclic monophosphate (cAMP) and several chemicals which elevate the intracellular level of cAMP on the induction of zygote formation in Micrasterias thomasiana var. notata. When added at a concentration of 0.5-3 mM, cAMP repressed the induction of zygotes and simultaneously promoted cell proliferation, although at a concentration of 0.1 mM it merely delayed the initiation of zygote induction. Methylxanthines caffeine (0.05-1 mM) and theophylline (0.05-1 mM), forskolin (10 mu M), which is a potent activator of adenylate cyclase, and a membrane-permeable cAMP analog, 8-bromo cAMP (0.1-3 mM), also repressed the induction of zygotes and simultaneously promoted cell proliferation. In contrast, another cAMP analog used in this study, N-6,O-2'-dibutyryl cAMP (2-3 mM), repressed the induction of zygotes but did not cause promotion of cell proliferation. This analog also specifically blocked the cell division directly involved with gamete formation. The results obtained suggest that intracellular cAMP may function as a signal which simultaneously represses zygote induction and causes proliferation of cells in Micrasterias.

 

DESNITSKI AG

On the origin of Metazoa
ZH OBSHCH BIOL 56 (5): 629-631 SEP-OCT 1995

Abstract:
Under consideration is a new version of the phagocytella conception. It is suggested to consider blastea not a particular organism but a morphologically variable taxon of mixotrophic flagellates. Some of its subtaxa might have made a transition to the true heterotrophy, thus having become direct metazoan ancestors.

 

WALTHER Z, HALL JL

The uni chromosome of chalmydomonashistone genes and nucleosome structure.
NUCLEIC ACIDS RES 23 (18): 3756-3763 SEP 25 1995

Abstract:
The uni linkage group (ULG) of Chlamydomonas reinhardtii contains many genes involved in the basal body-flagellar system, Recent evidence suggests that the corresponding uni chromosome is located in close proximity to the basal body complex. In the course of studies into its molecular organization, we have found a cluster of four histone genes on the ULG. The genes are arranged as divergently-transcribed pairs: H3-H4 and H2B-H2A. Genomic sequencing reveals that these genes lack introns and contain characteristic 3' palindromes similar to those of animals, The predicted amino acid sequences are highly conserved across species, with greatest similarities to the histone genes of Volvox. Southern analysis shows that each histone gene is present in 15-20 copies in Chlamydomonas and suggests a dispersed genomic organization. Northern analysis of mitotically-synchronized cells shows that, like the replication-dependent histones of higher eukaryotes, Chlamydomonas histone genes are expressed during S-phase. Using a gene-specific probe on Northern blots, we provide evidence that the ULG H4 gene is regulated in the same manner as other Chlamydomonas histone genes. Finally, micrococcal nuclease protection experiments show that the uni chromosome itself associates with histone proteins and displays a conventional nucleosomal banding pattern.

 

FABRY S, MULLER K, LINDAUER A, et al.

THE ORGANIZATION STRUCTURE AND REGULATORY ELEMENTS OF CHLAMYDOMONAS HISTONE GENES REVEAL FEATURES LINKING PLANT AND ANIMAL GENES
CURR GENET 28 (4): 333-345 SEP 1995

 

Abstract:
The genome of the green alga Chlamydomonas reinhardtii contains approximately 15 gene clusters of the nucleosomal (or core) histone H2A, H2B, H3 and H4 genes and at least one histone H1 gene. Seven non-allelic histone gene loci were isolated from a genomic library, physically mapped, and the nucleotide sequences of three isotypes of each core histone gene species and one linked H1 gene determined. The core histone genes are organized in clusters of H2A-H2B and H3-H4 pairs, in which each gene pair shows outwardly divergent transcription from a short (< 300 bp) intercistronic region. These intercistronic regions contain typically conserved promoter elements, namely a TATA-box and the three motifs TGGCCAGG(G/C)-CGAG, CGTTGACC and CGGTTG. Different from the genes of higher plants, but like those of animals and the related alga Volvox, the 3' untranslated regions contain no poly A signal, but a palindromic sequence (3' palindrome) essential for mRNA processing is present. One single H1 gene was found in close linkage to a H2A-H2B pair. The H1 upstream region contains the octameric promoter element GGTTGACC (also found upstream of the core histone genes) and two specific sequence motifs that are shared only with the Volvox H1 promoters. This suggests differential transcription of the H1 and the core histone genes. The H1 gene is interrupted by two introns. Unlike Volvox H3 genes, the three sequenced H3 isoforms are intron-free. Primer-directed PCR of genomic DNA demonstrated, however, that at least 8 of the about 15 H3 genes do contain one intron at a conserved position. In synchronized C. reinhardtii cells, H4 mRNA levels (representative of all core histone mRNAs) peak during cell division, suggesting strict replication-dependent gene control. The derived peptide sequences place C. reinhardtii core histones closer to plants than to animals, except that the H2A histones are more animal-like. The peptide sequence of histone H1 is closely related to the V. carteri VH1-II (66% identity). Organization of the core histone gene in pairs, and non-polyadenylation of mRNAs are features shared with animals, whereas peptide sequences and enhancer elements are shared with higher plants, assigning the volvocalean histone genes a position intermediate between animals and plants.

 

DESNITSKI AG

A REVIEW ON THE EVOLUTION OF DEVELOPMENT IN VOLVOX - MORPHOLOGICAL AND PHYSIOLOGICAL-ASPECTS
EUR J PROTISTOL 31 (3): 241-247 AUG 25 1995

Abstract:
This paper presents a morphophysiological concept of the evolution of Volvox development. We use published data concerning differences in size of the mature gonidia, rates of their division and peculiarities in segregation of somatic and reproductive cell lines. Based on this, four programmes (types) of asexual development of Volvox are recognized, and the evolutionary relationships among these programmes (but not among any concrete species of Volvox) are elucidated. The first developmental programme (Volvox powersii - V. pocockiae) is clearly primitive for the genus. This programme is characterized by ancestral features: large gonidia, division is fast, and there is no unequal (asymmetric) division into two cellular types. The second developmental programme(V. carteri), the third programme (V. tertius) and the fourth programme (V. aureus) are all derived, but constitute different phylogenetic trends. They each have some derived features: the second programme involves asymmetric division, the third programm involves slow division, while the fourth programme involves small gonidia and slow division. The evolutionary concept is supplemented by data on sexual reproduction in various species of Volvox.

 

GODL K, HALLMANN A, RAPPEL A, et al.

PHEROPHORINS - A FAMILY OF EXTRACELLULAR-MATRIX GLYCOPROTEINS FROM VOLVOX STRUCTURALLY RELATED TO THE SEX-INDUCING PHEROMONE
PLANTA 196 (4): 781-787 JUL 1995

Abstract:
Pherophorins are extracellular matrix (ECM) glycoproteins from Volvox that share homology with the sex-inducing pheromone. A novel pherophorin (pherophorin III) was characterized both with respect to expression pattern and proteolytic processing in vivo. Furthermore, it was shown that the pherophorins represent a protein family of ECM glycoproteins exhibiting a modular composition: their N-terminally located domain is a homolog of a domain found in the ECM glycoprotein SSG 185. Together with SSG 185, pherophorin I is a main component of the cellular zone within the ECM. The Volvox genome contains a tandem arrangement of genes encoding pherophorin II-related polypeptides. Inhibition of proteolytic processing of pherophorin II and III in vivo appears to result in the suppression of sexual induction.

 

DIETMAIER W, FABRY S, HUBER H, et al.

ANALYSIS OF A FAMILY OF YPT GENES AND THEIR PRODUCTS FROM CHLAMYDOMONAS-REINHARDTII
GENE 158 (1): 41-50 MAY 26 19

 

Abstract:
Small G-proteins encoded by the ras-like ypt genes are ubiquitous in eukaryotic cells. They have been shown to play an essential role in membrane vesicle transport. We have isolated four ypt genes, yptC1, yptC4, yptC5 and yptC6, from Chlamydomonas reinhardtii (Cr) genomic and cDNA libraries. Three of them, yptC1, yptC4 and yptC5, are close homologues of ypt genes previously found in the multicellular alga Volvox carteri (Vc), the fourth, yptC6, is new. Each yptC gene is present as a single copy in the genome. Comparisons of genomic and cDNA sequences revealed that the coding regions are interrupted by five (yptC5), six (yptC6), seven (yptC4) and eight (yptC1) introns, respectively. Cr ypt genes and the closely related Vc ypt genes have identical exon-intron structures, but the corresponding intron sequences are completely different. Polyadenylation is signalled by UAUAA, UGUAG and UGUAA. The deduced amino acid (aa) sequence of YptC6 exhibited 79% identity with HRab2; YptC1, YptC4 and YptC5 exhibited over 90% identity with their Vc homologues. Primary structures of the 9-aa 'effector domain' and the contiguous 'helix3-loop7' motif (approx. 30 aa) are 'diagnostic' features for functional assignment. Recombinant YptC proteins, overproduced in Escherichia coli and purified to near homogeneity, displayed strong and specific binding of GTP, but not of GMP or ATP. The four Cr Ypt proteins showed immunochemical cross reactions to their Vc counterparts. Moreover, Western blots demonstrated at least six types of Ypt in both Cr and Vc, suggesting that these Ypt are used for household functions responsible for vesicle transport rather than for cellular differentiation.

 

VOIGT J, VOGELER HP, KONIG WA, et al.

CHAOTROPE-SOLUBLE CELL-WALL GLYCOPROTEINS OF VOLVOX AND SOME MEMBERS OF THE ZYGNEMATOPHYCEAE IMMUNOLOGICALLY RELATED TO THE 150 KDA CELL-WALL GLYCOPROTEIN OF CHLAMYDOMONAS-REINHARDTII
MICROBIOL RES 150 (2): 129-137 MAY 1995

Abstract:
As previously described, the cell wall of Chlamydomonas reinhardtii contains several chaotrope-soluble glycoproteins with similar sugar compositions, but considerably different proportions of hydroxyproline. All these hydroxyproline-containing cell wall glycoproteins are recognized by a polyclonal antibody raised against the purified '150 kDa' cell wall glycoprotein of Chlamydomonas reinhardtii. This antibody also cross-reacts with some polypeptides present in the LiCl-extracts from intact cells of Spirotaenia erythrocephala, Spirotaenia obscura, Volvox aureus, and Volvox globator as revealed by Western blot analyses. Therefore, an Ige fraction of this particular antibody coupled to CNBr-activated Sepharose was used to isolate the immunologically related polypeptides extracted by aqueous LiCl from intact cells or cell wall preparations of different green algae belonging to the Volvocales or to the Zygnematophycene. Different and more or less complex polypeptide patterns were observed by comparative SDS-PAGE analyses of the polypeptide fractions isolated by immunoaffinity chromatography from extracts of Chlamydamonas reinhardtii, Volvox aureus, Micrasterias rotata, Gonatozygon brebissonii and Netrium digitus, respectively, whereas the corresponding fraction prepared from Spirotaenia erythrocephala contained a predominant '160 kDa' component. Amino acid and sugar analyses revealed that all these polypeptide fractions contained hydroxyproline (1.0-4.5 mol%) and the same sugars as the Chlamydomonas cell wall glycoproteins. However, the relative amounts of these sugars (arabinose, galactose, glucose, mannose and xylose) were found to be rather different. The highest proportion of hydroxyproline (4.5 mol%) and the highest ratio of arabinose:galactose (4.75:1) were determined for the glycoprotein fraction isolated from Spirotaenia erythrocephala indicating that at least the predominant '160 kDa' component is an intrinsic cell wall glycoprotein containing hydroxyproline and oligo-arabinosyl side chains. All the polypeptide fractions isolated from the other green algae obviously also contained chaotropesoluble cell wall glycoproteins as revealed by the presence of arabinose and hydroxyproline. These findings show that all the investigated green algae contain chaotrope-soluble cell wall glycoproteins immunologically related to the '150 kDa' cell wall glycoprotein of Chlamydomonas reinhardtii.

 

FABRY S, STEIGERWALD R, BERNKLAU C, et al.

STRUCTURE-FUNCTION ANALYSIS OF SMALL G-PROTEINS FROM VOLVOX AND CHLAMYDOMONAS BY COMPLEMENTATION OF SACCHAROMYCES-CEREVISIAE YPT/SEC MUTATIONS
MOL GEN GENET 247 (3): 265-274 MAY 10 1995

 

Abstract:
cDNAs representing nine small G protein genes encoding Ypt proteins from the green algae Volvox carteri (YptV) and Chlamydomonas reinhardtii (YptC) were tested for their ability to complement mutations in the YPT1, SEC4, and YPT7 genes of Saccharomyces cerevisiae strains defective in different steps of intracellular vesicle transport. None of the heterologously expressed algal genes was able to complement mutations in SEC4 or YPT7, but three of them, yptV1, yptC1, and yptV2, restored a YPT1 null mutation. On the amino acid sequence level, and particularly with respect to known small G protein specificity domains. YptV1p and YptC1p are the closest algal analogs of yeast Ypt1p, with 70% overall identity and identical effector regions, but YptV2p is only 55% identical to Ypt1p, and its effector domain resembles that of Sec4p. To define more precisely the regions that supply Ypt1p function, six chimeras were constructed by reciprocal exchange of 68/72-, 122/123-, and 162/163-amino acid segments of the C-terminal regions between YptV1p (complementing) and YptV3p (non-complementing). Segments containing 68 amino acids of the hypervariable C-terminal, and 41 residues of the N-terminal region including the effector region, of YptV1p could be replaced by the corresponding parts of YptV3p without loss of function in yeast, but exchanges within the central core destroyed the ability to rescue the YPT1 mutation. Sequence analysis of ypt1-complementing and -noncomplementing Ypt types suggests that surface loop3 represents a novel specificity domain of small G proteins.

 

FELDWISCH O, LAMMERTZ M, HARTMANN E, et al.

PURIFICATION AND CHARACTERIZATION OF A CAMP-BINDING PROTEIN OF VOLVOX-CARTERI F NAGARIENSIS IYENGAR
EUR J BIOCHEM 228 (2): 480-489 MAR 1 1995

Abstract:
Two cAMP-binding proteins, cbp1 and cbp2, were purified from the cytoplasm of the green alga Volvox carteri. Both proteins have a native molecular mass of 90 kDa as determined by gel filtration. cbp2 was purified to apparent electrophoretic homogeneity, having a subunit molecular mass of 42 kDa as determined by SDS/PAGE. The cbp1 preparation contains a 42-kDa and a 44-kDa band. The cAMP-binding activity is not associated with protein kinase activity. Tryptic peptides of cbp2 were sequenced by automated Edman degradation. Two pairs of peptides differ in one amino acid only, thus pointing to the presence of isoforms of cbp2. Both binding proteins differed from the cAMP-specific phosphodiesterases of V. carteri with respect to charge, molecular mass and binding affinity to N-6-cAMP-agarose, Reverse-phase chromatography of the bound ligand revealed that the two binding proteins hydrolyse cAMP to 5'AMP. The binding specificity of purified cbp1 and cbp2 was probed by a set of modified cAMP derivatives. Both proteins bind cAMP strictly specifically in the anti conformation; position 1 and 6 of the adenine moiety and at least one of the exocyclic O atoms of the ribose cyclic phosphate moiety are essential. 3-Isobutyl-1-methylxanthine is an effective inhibitor of binding but the natural methylxyanthines are not. At present it is not clear whether cbp1 and cbp2 are individual proteins or isoforms of one another.

 

 

MILLER SM, KIRK DL

CHARACTERIZATION OF A VOLVOX GENE REGULATING ASYMMETRIC CELL-DIVISION
MOL BIOL CELL 6: 501-501 Suppl. S NOV 1995