Construction of a gamete-enriched gene pool and RNAi-mediated functional analysis in Dictyostelium discoideum.
Macrocysts in Dictyostelium discoideum possess prototypic features of sexual reproduction and are useful for understanding the basic mechanisms of the reproductive process. Here, we randomly analyzed 1,071 gamete cDNAs, and then constructed a gamete-specific subtraction library, FC-IC. Nucleotide sequences of all 903 FC-IC clones were determined and clustered into 272 independent genes. Expression analysis based on real-time RT-PCR revealed 67 gamete-enriched genes, among which those involved in 'signal transduction' and 'multicellular organization' are prevalent. One of them, FC-IC0003, appeared also to be mating-type specific, and was named gmsA. RNAi-mediated silencing as well as disruption of gmsA reduced the cellular competency for sexual cell fusion, indicating the involvement of this gene in the sexual development of D. discoideum.
Muramoto
Tetsuya
T
Institute of Biological Sciences, University of Tsukuba, Tsukuba-shi 305-8572, Japan.
Suzuki
Katsuya
K
Shimizu
Hajime
H
Kohara
Yuji
Y
Kohriki
Eiko
E
Obara
Shinji
S
Tanaka
Yoshimasa
Y
Urushihara
Hideko
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004023:Dictyostelium / Q000235:genetics*; D020869:Gene Expression Profiling; D015723:Gene Library; D005788:Gene Pool; D005854:Germ Cells / Q000502:physiology*; D008969:Molecular Sequence Data; D016372:RNA, Antisense / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering; D016415:Sequence Alignment; D015398:Signal Transduction
D016372:RNA, Antisense; D034741:RNA, Small Interfering