Molecular cloning and preliminary functional analysis of two novel human KRAB zinc finger proteins, HKr18 and HKr19.
cDNA clones encoding two novel human KRAB zinc finger proteins, HKr18 and HKr19, were isolated from a human testis cDNA library. Their corresponding genes were later identified in sequences originating from chromosomes 19 and 7, respectively. On the basis of the collected information from gene and cDNA sequences, Hkr18 was found to be a protein of 94 kDa with 20 zinc finger motifs in its C terminus. The HKr19 is a smaller protein, with a molecular weight of 56 kDa containing 11 zinc finger motifs. Both HKr18 and HKr19 contained a KRAB A as well as a KRAB B domain in their N termini. Northern blot analysis showed expression of HKr18 in all human tissues tested, indicating a ubiquitous expression pattern. In contrast, HKr19 showed a more restricted tissue distribution, with detectable expression primarily in testis and fetal tissues. The HKr19 protein is a member of the large ZNF91 subfamily of KRAB zinc finger genes. A PCR-based analysis of the expression of HKr19 and other closely related genes showed that lymphoid, myeloid, and nonhematopoietic cells expressed different sets of these genes. This latter finding indicates that some members of the ZNF91 family may be involved in regulating lineage commitment during hematopoietic development. Transfection of various parts of HKr19 into human embryonic kidney cells (HEK 293 cells) showed that the entire protein and its zinc finger region were toxic to these cells when expressed at high levels. In contrast, the KRAB domain and the linker region seemed to be well tolerated.
Mark
C
C
Department of Cell and Molecular Biology, Biomedical Center, Uppsala, Sweden.
Looman
C
C
Abrink
M
M
Hellman
L
L
D000328:Adult; D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D002460:Cell Line; D003001:Cloning, Molecular; D016895:Culture Media, Serum-Free; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005333:Fetus / Q000502:physiology; D015723:Gene Library; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D012097:Repressor Proteins; D013737:Testis / Q000737:chemistry* / Q000502:physiology; D014162:Transfection
D016895:Culture Media, Serum-Free; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D012097:Repressor Proteins; C422407:ZNF350 protein, human