Differential gene expression in CD3epsilon- and RAG1-deficient thymuses: definition of a set of genes potentially involved in thymocyte maturation.
A set of 3000 mouse thymus cDNAs was analyzed by extensive measurement of expression using complex-probe hybridization of DNA arrays ("quantitative differential screening"). The complex probes were initially prepared using total thymus RNA isolated from C57BL/6 wild-type (WT), CD3epsilon- and RAG1-deficient mice. Over 100 clones displaying over- or under-expression by at least a factor of two between WT and knockout (KO) thymuses were further analyzed by measuring hybridization signatures with probes from a wide range of KO thymuses, cell types, organs, and embryonic thymuses. A restricted set of clones was selected by virtue of their expression spectra (modulation in KO thymuses and thymocytes, lymphoid cell specificity, and differential expression during embryonic thymus development), sequenced at one extremity, and compared to sequences in databases. Clones corresponding to previously identified genes (e.g., Tcrbeta, Tcf1 or CD25) showed expression patterns that were consistent with existing data. Ten distinct clones corresponding to new genes were subjected to further study: Northern blot hybridization, in situ hybridization on thymus sections, and partial or complete mRNA sequence determination. Among these genes, we report a new serine peptidase highly expressed in cortical epithelial cells that we have named thymus-specific serine peptidase (TSSP), and an acidic protein expressed in thymocytes and of unknown function that we have named thymus-expressed acidic protein (TEAP). This approach identifies new molecules likely to be involved in thymocyte differentiation and function.
Carrier
A
A
Centre d'Immunologie INSERM-CNRS de Marseille-Luminy, Case 906, Marseille, France. carrier@ciml.univ-mrs.fr
Nguyen
C
C
Victorero
G
G
Granjeaud
S
S
Rocha
D
D
Bernard
K
K
Miazek
A
A
Ferrier
P
P
Malissen
M
M
Naquet
P
P
Malissen
B
B
Jordan
B R
BR
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D017252:CD3 Complex / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D005786:Gene Expression Regulation; D019672:Genes, T-Cell Receptor / Q000235:genetics; D018398:Homeodomain Proteins / Q000378:metabolism*; D006801:Humans; D017403:In Situ Hybridization; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000378:metabolism; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013950:Thymus Gland / Q000254:growth & development* / Q000378:metabolism; D014018:Tissue Distribution
D017252:CD3 Complex; C121704:CD3 antigen, zeta chain; C115335:CD3E protein, human; D018076:DNA, Complementary; D018398:Homeodomain Proteins; D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell; C064658:RAG-1 protein