Molecular cloning of a novel sex pheromone responsible for the release of a different sex pheromone in Closterium peracerosum-strigosum-littorale complex.
A sex pheromone, protoplast-release-inducing protein (PR-IP) inducer, of the Closterium peracerosum-strigosum-littorale complex is known to induce the release of PR-IP, from mating-type plus (mt+) cells during sexual reproduction. The purified PR-IP inducer was treated with trypsin to obtain internal peptides for determination of partial amino acid sequences. Using these sequences, oligonucleotides were synthesized and used as primers for the combined reverse transcription-PCR. A 296 bp cDNA fragment was amplified, permitting the cloning of corresponding full length cDNA (CpPI; Closterium peracerosum-strigosum-littorale complex PR-IP inducer). The deduced amino acid sequence of CpPI encodes a protein of 212 amino acid residues of M(r) 23,071 whereas portion of the peptide secreted is predicted to have 142 amino acid residues of M(r) 15,717 and shows no significant similarity with known proteins. The predicted protein has three possible consensus sequences for asparagine-linked glycosylation site. The CpPI gene was expressed when mating-type minus (mt-) cells were incubated at a low cell density in the light. Nitrogen deprivation from the medium enhances expression of the CpPI gene. An analysis by genomic Southern hybridization revealed that the cDNA probe hybridized to several DNA fragments obtained from both the genome of mt- and mt+ cells. However, in mt- cells, transcripts for the PR-IP inducer could not be detected by Northern hybridization.
Sekimoto
H
H
Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Japan. sekimoto@ims.u-tokyo.ac.jp
Fukumoto
R
R
Dohmae
N
N
Takio
K
K
Fujii
T
T
Kamiya
Y
Y
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003227:Conjugation, Genetic; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D018521:Plant Physiological Phenomena; D010944:Plants / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012724:Sex Attractants / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*
D010446:Peptide Fragments; D011994:Recombinant Proteins; D012724:Sex Attractants