Molecular cloning and characterization of chemokine-like factor super family member 1 (CKLFSF1), a novel human gene with at least 23 alternative splicing isoforms in testis tissue.
Chemokine-like factor (CKLF) was isolated from PHA-stimulated U937 cells. It is composed of 152 amino acids and located on chromosome 16q22. Utilizing bioinformatics, based on CKLF cDNA and protein sequences, in combination with experimental validation, we identified a novel gene designated chemokine-like factor super family member 1 (CKLFSF1). CKLFSF1 maps on chromosome 16q22, and the full-length gene comprises of seven exons and six introns. Using RACE-PCR, we identified two potential alternative transcription start sites, 1A and 1B. Northern blot and RT-PCR analysis demonstrated that CKLFSF1 is predominantly expressed in human testis tissue, with only lower levels of expression in many other human tissues. RT-PCR and cDNA sequencing identified 23 alternatively spliced isoforms of CKLFSF1 in the testis tissue, which encode protein variants ranging from 36 to 169 amino acids in length. Immunohistochemistry analysis demonstrated that CKLFSF1 proteins are highly expressed in spermatocyte and in tissue fluid of human testes tissue. In light of these findings, we propose that CKLFSF1 may play an important role in spermatogenesis or testicular development.
Wang
Lu
L
Peking University Center for Human Disease Genomics, 38 Xueyuan Road, Beijing 100083, China.
Wu
Chunxiao
C
Zheng
Ying
Y
Qiu
Xiaoyan
X
Wang
Li
L
Fan
Hui
H
Han
Wenling
W
Lv
Bingfeng
B
Wang
Ying
Y
Zhu
Xiaohui
X
Xu
Mingxu
M
Ding
Peiguo
P
Cheng
Shanhong
S
Zhang
Yingmei
Y
Song
Quansheng
Q
Ma
Dalong
D
D017398:Alternative Splicing; D001483:Base Sequence; D018925:Chemokines / Q000032:analysis / Q000235:genetics* / Q000502:physiology; D002885:Chromosomes, Human, Pair 16; D003001:Cloning, Molecular; D040461:Gene Components; D006801:Humans; D062792:MARVEL Domain-Containing Proteins; D008297:Male; D008969:Molecular Sequence Data; D020033:Protein Isoforms; D012333:RNA, Messenger / Q000235:genetics; D013737:Testis / Q000737:chemistry* / Q000254:growth & development; D014018:Tissue Distribution; D024363:Transcription Initiation Site; D020298:U937 Cells
C432676:CKLF protein, human; D018925:Chemokines; D062792:MARVEL Domain-Containing Proteins; D020033:Protein Isoforms; D012333:RNA, Messenger