ALG2, the Hansenula polymorpha isocitrate lyase gene.
To set the basis for molecular and cellular studies of the glyoxylate cycle in methylotrophic yeasts, we isolated and characterized ALG2, the Hansenula polymorpha isocitrate lyase gene. Complementation work and sequence analysis revealed an ORF of 1458 nucleotides, encoding a 486 amino acid protein with a predicted molecular mass of 54.9 kDa. This protein is shorter than the Saccharomyces cerevisiae and Candida tropicalis ICLs, lacks a PST1 signal and possesses a PTS2-like signal. The transcriptional regulation of ALG2 mRNA levels by carbon source is mainly achieved by glucose repression-derepression, whereas ethanol induction plays only a minor role. We present evidence indicating that, in H. polymorpha, neither isocitrate lyase activity nor the ALG2 gene product are necessary for C(1)-peroxisome degradation triggered by ethanol. Therefore, the involvement of glyoxylate in degradation, as described by Kulachkovsky et al. (1997) for Pichia methanolica, does not necessarily apply to all methylotrophic yeasts. The relevant nucleotide sequence has been deposited at GenBank (Accession No. AF373067.1).
Berardi
Enrico
E
Laboratorio di Genetica Microbica, Dipartimento di Biotecnologie, Università Politecnica delle Marche, Via Brecce Bianche, 60131 Ancona, Italy. berardi@univpm.it
Gambini
Annalisa
A
Bellu
Anna Rita
AR
D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D005816:Genetic Complementation Test; D007522:Isocitrate Lyase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D020675:Peroxisomes / Q000378:metabolism; D010843:Pichia / Q000201:enzymology* / Q000235:genetics*; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D016415:Sequence Alignment; D014158:Transcription, Genetic
D012331:RNA, Fungal; D012333:RNA, Messenger; D007522:Isocitrate Lyase