Primary structure of the plasma membrane H+-ATPase from the halotolerant alga Dunaliella bioculata

Abstract

P-type ATPase-specific oligodeoxyribonucleotides were used to obtain a fragment of the H(+)-ATPase of the salt tolerant alga Dunaliella bioculata by polymerase chain reaction (PCR). This fragment served as a probe in screening a cDNA-library from this organism. The complete primary structure of the ATPase protein (DBPMA1) was deduced from sequencing a 4.7 kb cDNA clone. The protein shows highest homology to H(+)-ATPases from higher plants and fungi (43% identity, 67% similarity) but has a higher calculated molecular mass (123 kDa). The latter can be assigned mainly to an additional hydrophilic domain between transmembrane segments VI and VII and to an extended carboxyterminus. These unusual structural features of DBPMA1 are interpreted in terms of providing regulatory sites of the enzyme. Southern blot analysis suggests the presence of only a single copy of the gene in the haploid D. bioculata genome. To investigate the role of the H(+)-ATPase in the adaption of D. bioculata to different external NaCl concentrations, we employed northern blot analyses. The results indicate that the pma1 transcript level of cells growing in salinities between 0.1 and 3 M NaCl is not directly correlated with the external salt concentration.

Influence of photoheterotrophy on the expression of chlorophyll a/b-binding proteins in the green alga Pyrobotrys stellata

“…The codon usage of both cab-genes is strongly biased towards codons ending in C or G, as found for other highly expressed chlorophycean genes (Siflow et al 1985;Goldschmidt-Glermont and Rahire 1986;LaRoche et al 1990;Wolf et al 1993Wolf et al , 1995. The putative polyadenylation signal of green algae, TGTAA (Siflow et al 1985), which is also present in other algal cab-genes (Imbault et al 1988;Long et al 1989;LaRoche et al 1990;Larouche et al 1991) was found in both transcripts (Figures 1 and 2).…”

Molecular cloning of a P-type Ca2+-ATPase from the halotolerant alga Dunaliella bioculata

“…We previously reported on the cloning of the PM H+-ATPas6 (DBPMA1) from D. bioculata (Wolf et al 1995). We previously reported on the cloning of the PM H+-ATPas6 (DBPMA1) from D. bioculata (Wolf et al 1995).…”

“…Also, no sequence similarities to any of the known CaM-binding sites in different enzymes, including those that slightly deviat} from the general scheme, were detected. Contrarily, major rearrangements occurred with the P M H § in D. bioculata (Wolf et al 1995). I L EQF D D MENAHTKTVEEVLAYFGVNESTGL S LEQVKKLKEKWGSNELPAEEGKTLLELVI EQFED~~ MENAHTKTVEEVLGHFGVNE STGL SLEQVKKLKERWGSNELPAEEGKTLLELVI EQFEDD~I~ TMI -SEGESI---Q ERNAE~ I EQLKS YEADDATVLRNGQ --LQLI P SADI VPGDIVE LAVGNKVPADTRVSH I YTTSL K VNQDETGESGF ESNAEKALEALKEMQGESAKVLRDGYL -VPDF PAKELV PGDIVELRVGDKVPADMRVATLKS STLR FEEGEET---I ERNAENA I EALKEYE PI~GKVYRADR KAVQRI KARDLVPGDI AEVAVGDKVPADIRI I S IKSTTLR FE EHDDEAEQL EKNAESAIEALKEYEPEMGKVIRADKTGIQKI KARDLVPGDI VE I SVGDKI PADLRLI S I Fig.…”

“…Genomic DNA was isolated, fragmented with restriction endonucleases, and used for Southern analyses as described previously (Wolf et al 1995). For RNA blot hybridization analysis, approx.…”

Mechanisms of Acido-Tolerance and Characteristics of Photosystems in an Acidophilic and Thermophilic Red Alga, Cyanidium Caldarium

“…The deduced amino acid sequence was more homologous to those of plasma membrane H + -ATPases from higher plants (86.5% similarity and 54.1% identity) than to that from the salt-tolerant green alga Dunaliella bioculata (75.1% similarity and 40.1% identity). The C-terminal region of plasma membrane H + -ATPase from higher plants has been reported to serve as an autoinhibitory domain, important for regulation of its activity (Wolf et al, 1995). A homologous sequence, corresponding to the autoinhibitory domain, however, was not found in the C-terminal region of the plasma membrane H + -ATPase from Cyanidium.…”