Identification and characterization of three differentially expressed genes, encoding S-adenosylhomocysteine hydrolase, methionine aminopeptidase, and a histone-like protein, in the toxic dinoflagellate Alexandrium fundyense.
Genes showing differential expression related to the early G(1) phase of the cell cycle during synchronized circadian growth of the toxic dinoflagellate Alexandrium fundyense were identified and characterized by differential display (DD). The determination in our previous work that toxin production in Alexandrium is relegated to a narrow time frame in early G(1) led to the hypothesis that transcriptionally up- or downregulated genes during this subphase of the cell cycle might be related to toxin biosynthesis. Three genes, encoding S-adenosylhomocysteine hydrolase (Sahh), methionine aminopeptidase (Map), and a histone-like protein (HAf), were isolated. Sahh was downregulated, while Map and HAf were upregulated, during the early G(1) phase of the cell cycle. Sahh and Map encoded amino acid sequences with about 90 and 70% similarity to those encoded by several eukaryotic and prokaryotic Sahh and Map genes, respectively. The partial Map sequence also contained three cobalt binding motifs characteristic of all Map genes. HAf encoded an amino acid sequence with 60% similarity to those of two histone-like proteins from the dinoflagellate Crypthecodinium cohnii Biecheler. This study documents the potential of applying DD to the identification of genes that are related to physiological processes or cell cycle events in phytoplankton under conditions where small sample volumes represent an experimental constraint. The identification of an additional 21 genes with various cell cycle-related DD patterns also provides evidence for the importance of pretranslational or transcriptional regulation in dinoflagellates, contrary to previous reports suggesting the possibility that translational mechanisms are the primary means of circadian regulation in this group of organisms.
Taroncher-Oldenburg
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Biology Department, Woods Hole Oceanographic Institution, Woods Hole, Massachusetts 02543, USA. gto@mit.edu
Anderson
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D043383:Adenosylhomocysteinase; D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000737:chemistry / Q000235:genetics*; D000818:Animals; D002453:Cell Cycle; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics; D004141:Dinoflagellida / Q000166:cytology / Q000201:enzymology / Q000235:genetics*; D029968:Escherichia coli Proteins; D005786:Gene Expression Regulation; D017125:Genes, Protozoan; D006867:Hydrolases / Q000737:chemistry / Q000235:genetics*; D063208:Methionyl Aminopeptidases; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014118:Toxins, Biological / Q000096:biosynthesis*
D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D014118:Toxins, Biological; C116822:hns protein, E coli; D006867:Hydrolases; D043383:Adenosylhomocysteinase; D000626:Aminopeptidases; D063208:Methionyl Aminopeptidases