Identification and characterization of a RING zinc finger gene (C-RZF) expressed in chicken embryo cells.
To identify changes in gene expression that occur in chicken embryo brain (CEB) cells as a consequence of their binding to the extracellular matrix molecule cytotactin/tenascin (CT/TN), a subtractive hybridization cloning strategy was employed. One of the cDNA clones identified was predicted to encode 381 amino acids and although it did not resemble any known sequences in the nucleic acid or protein data bases, it did contain the sequence motif for the cysteine-rich C3HC4 type of zinc finger, also known as a RING-finger. This sequence was therefore designated the chicken-RING zinc finger (C-RZF). In addition to the RING-finger, the C-RZF sequence also contained motifs for a leucine zipper, a nuclear localization signal, and a stretch of acidic amino acids similar to the activation domains of some transcription factors. Southern analysis suggested that C-RZF is encoded by a single gene. Northern and in situ hybridization analyses of E8 chicken embryo tissues indicated that expression of the C-RZF gene was restricted primarily to brain and heart. Western analysis of the nuclear and cytoplasmic fractions of chicken embryo heart cells and immunofluorescent staining of chicken embryo cardiocytes with anti-C-RZF antibodies demonstrated that the C-RZF protein was present in the nucleus. The data suggest that we have identified another member of the RING-finger family of proteins whose expression in CEB cells may be affected by CT/TN and whose nuclear localization and sequence motifs predict a DNA-binding function in the nucleus.
Tranque
P
P
Department of Neurobiology, The Scripps Research Institute, La Jolla, CA 92037, USA.
Crossin
K L
KL
Cirelli
C
C
Edelman
G M
GM
Mauro
V P
VP
D000595:Amino Acid Sequence; D000818:Animals; D030161:Avian Proteins; D001483:Base Sequence; D001921:Brain / Q000196:embryology / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D002642:Chick Embryo; D017124:Conserved Sequence; D003593:Cytoplasm / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000737:chemistry; D015870:Gene Expression; D006321:Heart / Q000196:embryology; D017403:In Situ Hybridization; D016350:Leucine Zippers; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism*; D009687:Nuclear Proteins; D016366:Open Reading Frames; D015347:RNA Probes; D017386:Sequence Homology, Amino Acid; D014981:Xenopus; D016335:Zinc Fingers
D030161:Avian Proteins; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; C099027:RING finger protein 13, chicken; D015347:RNA Probes