서지주요정보
Identification and characterization of GATA factor homologs in schizosaccharomyces pombe = 분열 효모에서 GATA 조절 유사인자의 분리 및 분석
서명 / 저자 Identification and characterization of GATA factor homologs in schizosaccharomyces pombe = 분열 효모에서 GATA 조절 유사인자의 분리 및 분석 / Kwang-Lae Hoe.
저자명 Hoe, Kwang-Lae ; 허광래
발행사항 [대전 : 한국과학기술원, 1996].
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8006634

소장위치/청구기호

학술문화관(문화관) 보존서고

DBS 96001

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초록정보

GATA-binding proteins constitute one of transcription factor families, which bind to the consensus target site "WGATAR" (W=A or T and R=A or G). GATA factors, whose distinct feature is the zinc finger motif $Cys-X_2-Cys-X_{17}-Cys-X_2-Cys$, are present in many eukaryotes from yeast to human. GATA factors of vertebrates are shown to be involved in the regulation of globin gene expression or other developmentally related genes. On the other hand in fungi, GATA factors regulate expression of genes involved in the regulation of nitrogen or iron metabolism. As an attempt toward elucidation for the functions of GATA factors in Shizosaccharomyces pombe, we have isolated and characterized the two GATA factor homologous genes, gaf1 and gaf2. Genomic Southern analysis, with the degenerate oligo probe encoding the well-conserved zinc finger core region, showed the possibility of the presence of at least 2 GATA factor homologs in S. pombe. The gaf1 gene was first cloned by the method of PCR with degenerate primers corresponding to the conserved sites in the zinc finger region. The second GATA factor gene, gaf2, was cloned by the low stringent screening of a λ-S. pombe genomic library using the partial gaf1 gene as a probe. Analysis of the DNA sequences showed that Gaf1 and Gaf2 have the 290- and 564-amino acid ORF, respectively. Northern analysis showed that the sizes of gaf1, gaf2 transcripts are 4.2 kb with 2 minor bands (1.6 and 2.1 kb) and 3.1 kb with a minor band at 1.6 kb, respectively. The transcription levels of gaf1 and gaf2 were not affected by a nitrogen source in media. Disruption of gaf1 was not lethal and showed no detectable change in the growth rate. The gel mobility shift assay, using the GST-Gaf1 fusion protein and the canonical GATA motif (the DAL7 UAS of S. cerevisiae), showed that Gaf1 binds to GATA motifs with specificity. In addition, the transactivation assay, where the lacZ expression was examined under the GAL4-CYC1 hybrid promoter, showed that Gaf1 functions as a transcription activator. As a consequence, It is certain that Gaf1 is a new member of the GATA family which can function as a transcriptional activator of its presumptive target genes. The function of gaf2 is not known yet, but there is an indication that it might be involved in meiosis pathway of S. pombe.

서지기타정보

서지기타정보
청구기호 {DBS 96001
형태사항 viii, 110 p. : 삽도 ; 26 cm
언어 영어
일반주기 저자명의 한글표기 : 허광래
지도교수의 영문표기 : Ook-Joon Yoo
지도교수의 한글표기 : 유욱준
학위논문 학위논문(박사) - 한국과학기술원 : 생물과학과,
서지주기 Reference : p. 98-104
주제 GATA Factor
PCR
Zinc Finger
Transcriptional Activator
가타 인자
연쇄 중합 반응
아연 손
전사 활성 인자
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