Characterization of the rat gamma-crystallin gene family and its expression in the eye lens.
Rat genomic clones, which together contain all of the rat genomic gamma-crystallin sequences, have been characterized. Five gamma-crystallin genes are located on a contiguous DNA region, 63 X 10(3) base-pairs long. These genes, named (5') gamma 1-1, gamma 1-2, gamma 2-2 and gamma 3-1 (3'), are all oriented head to tail. A sixth gamma-crystallin gene, named the gamma 4-1 gene, could not be linked to the gamma-crystallin gene cluster with our present set of genomic clones. Mapping experiments using single copy sequences which form the extreme 5' or 3' region of the gene cluster showed that, if the gamma 4-1 gene is located on the same chromosome, then it must be separated from the gene cluster by at least 25 X 10(3) base-pairs of DNA. All gamma-crystallin genes have a similar mosaic structure. They contain a large (0.9 X 10(3) to 1.88 X 10(3) base-pairs) intron in the middle of the gene and are further interrupted close to the 5' end of the gene. The length of the first exon varies from about 40 to about 50 base-pairs. The complementary DNA clone pRL-gamma-3 used in this study is a copy of the transcript of the gamma 3-1 gene, while the second complementary DNA clone, pRL-gamma-2, is most likely a copy of the transcript of the gamma 2-1 gene. It is further shown that rat lens messenger RNA protects fragments from the 3' ends of the four other gamma-crystallin genes against degradation by S1 nuclease, hence all six gamma-crystallin genes present in the rat genome must be transcribed in the lens. Repetitive sequences were found to be present between and around the gamma-crystallin genes. Mapping with cloned repetitive sequences showed that three different repeats, designated A, B and C, occur more than once in the gamma-crystallin gene cluster. Repeat C is also found in the gamma 4-1 region. A repetitive region 3' to the gamma 3-1 gene contains members of all three repeat families.
Moormann
R J
RJ
den Dunnen
J T
JT
Heuyerjans
J
J
Jongbloed
R J
RJ
van Leen
R W
RW
Lubsen
N H
NH
Schoenmakers
J G
JG
D000818:Animals; D001345:Autoradiography; D001483:Base Sequence; D003001:Cloning, Molecular; D003459:Crystallins / Q000235:genetics*; D004247:DNA; D004262:DNA Restriction Enzymes; D005796:Genes; D008040:Genetic Linkage; D007908:Lens, Crystalline / Q000032:analysis*; D009693:Nucleic Acid Hybridization; D051381:Rats; D012091:Repetitive Sequences, Nucleic Acid; D014158:Transcription, Genetic
D003459:Crystallins; D004247:DNA; D004262:DNA Restriction Enzymes