Isolation and characterization of a single-copy actin gene from a sterile mutant of Ulva pertusa (Ulvales, Chlorophyta).
We constructed a cDNA library from sterile Ulva pertusa (Ulvales, Chlorophyta), and isolated and characterized a full-length cDNA clone encoding actin. The actin (ACT) cDNA consisted of 1487 nucleotides (nt) and had an open reading frame (ORF) encoding a polypeptide of 377 amino acid (AA) residues. The ACT gene had one intron in the 5'-untranslated region and three introns in the coding region. Transcription started 26 nt downstream of the putative TATA box. A potential polyadenylation signal, TGTAG, was located 100 nt downstream of the terminator codon, TAG. Amino acid alignment with actins from various algae and land plants showed that sterile U. pertusa actin was more similar to actins from Chlorophyta, Phaeophyta, Euglenophyta, and higher plants (over 76.9%) than to actins from Rhodophyta. Southern blot analysis indicated that the sterile U. pertusa genome has only a single actin-encoding gene. Thalli grown on a 12D/12L photoperiod increased in surface area some two-fold over 24 h regardless of the nutritional conditions. The growth rate of thalli during the light period was significantly higher than that during the dark period. Northern hybridization indicated that the expression of actin mRNA was induced and repressed by the light and dark treatments, respectively. These results suggest that the U. pertusa cell division cycle has a periodicity of 24 h and that the ACT gene is highly transcribed during cell growth and development in the light period.
Kakinuma
Makoto
M
Laboratory of Marine Biochemistry, Faculty of Bioresources, Mie University, Tsu Mie 514-8507, Japan.
Coury
Daniel A
DA
Inagaki
Eri
E
Itoh
Sayaka
S
Yoshiura
Yasutoshi
Y
Amano
Hideomi
H
D000199:Actins / Q000235:genetics*; D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D003470:Culture Media / Q000494:pharmacology; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D018628:Gene Dosage; D015870:Gene Expression / Q000187:drug effects; D005796:Genes / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D017440:Photoperiod; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D044445:Ulva / Q000235:genetics* / Q000254:growth & development / Q000528:radiation effects
D000199:Actins; D003470:Culture Media; D012333:RNA, Messenger; D004247:DNA