PUBLICATION
A novel puf-A gene predicted from evolutionary analysis is involved in the development of eyes and primordial germ-cells
- Authors
- Kuo, M.W., Wang, S.H., Chang, J.C., Chang, C.H., Huang, L.J., Lin, H.H., Yu, A.L., Li, W.H., and Yu, J.
- ID
- ZDB-PUB-090330-9
- Date
- 2009
- Source
- PLoS One 4(3): e4980 (Journal)
- Registered Authors
- Keywords
- Zebrafish, Eyes, Embryos, Small interfering RNAs, Multiple alignment calculation, Untranslated regions, Ovaries, In situ hybridization
- MeSH Terms
-
- Animals
- Biological Evolution
- Cell Lineage
- Cell Movement
- Cloning, Molecular
- DNA-Binding Proteins/analysis
- DNA-Binding Proteins/physiology*
- Eye/embryology
- Eye/growth & development*
- Germ Cells/growth & development*
- Humans
- Mice
- RNA, Messenger
- RNA-Binding Proteins/analysis
- RNA-Binding Proteins/physiology*
- Zebrafish
- Zebrafish Proteins/analysis
- Zebrafish Proteins/physiology*
- PubMed
- 19319195 Full text @ PLoS One
Citation
Kuo, M.W., Wang, S.H., Chang, J.C., Chang, C.H., Huang, L.J., Lin, H.H., Yu, A.L., Li, W.H., and Yu, J. (2009) A novel puf-A gene predicted from evolutionary analysis is involved in the development of eyes and primordial germ-cells. PLoS One. 4(3):e4980.
Abstract
Although the human genome project has been completed for some time, the issue of the number of transcribed genes with identifiable biological functions remains unresolved. We used zebrafish as a model organism to study the functions of Ka/Ks-predicted novel human exons, which were identified from a comparative evolutionary genomics analysis.In this study, a novel gene, designated as puf-A, was cloned and functionally characterized, and its homologs in zebrafish, mouse, and human were identified as one of the three homolog clusters which were consisted of 14 related proteins with Puf repeats. Computer modeling of human Puf-A structure and a pull-down assay for interactions with RNA targets predicted that it was a RNA-binding protein. Specifically, Puf-A contained a special six Puf-repeat domain, which constituted a unique superhelix half doughnut-shaped Puf domain with a topology similar to, but different from the conventional eight-repeat Pumilio domain. Puf-A transcripts were uniformly distributed in early embryos, but became restricted primarily to eyes and ovaries at a later stage of development. In mice, puf-A expression was detected primarily in retinal ganglion and pigmented cells. Knockdown of puf-A in zebrafish embryos resulted in microphthalmia, a small head, and abnormal primordial germ-cell (PGC) migration. The latter was confirmed by microinjecting into embryos puf-A siRNA containing nanos 3' UTR that expressed in PGC only. The importance of Puf-A in the maturation of germline stem cells was also implicated by its unique expression in the most primitive follicles (stage I) in adult ovaries, followed by a sharp decline of expression in later stages of folliculogenesis. Taken together, our study shows that puf-A plays an important role not only in eye development, but also in PGC migration and the specification of germ cell lineage. These studies represent an exemplary implementation of a unique platform to uncover unknown function(s) of human genes and their roles in development regulation.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping