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T Honjo

Publications and source records attributed to T Honjo.

At least 379 records · Page 21Linked to original sources
PubMed1982

The nucleotide sequence of the mouse immunoglobulin epsilon gene: comparison with the human epsilon gene sequence.

We have determined the nucleotide sequence of the immunoglobulin epsilon gene cloned from newborn mouse DNA. The epsilon gene sequence allows prediction of the amino acid sequence of the constant region of the epsilon chain and comparison of it with sequences of the human epsilon and other mouse immunoglobulin genes. The epsilon gene was shown to be under the weakest selection pressure at the protein level among the immunoglobulin genes although the divergence at the synonymous position is similar. Our results suggest that the epsilon gene may be dispensable, which is in accord with the fact that IgE has only obscure roles in the immune defense system but has an undesirable role as a mediator of hypersensitivity. The sequence data suggest that the human and murine epsilon genes were derived from different ancestors duplicated a long time ago. The amino acid sequence of the epsilon chain is more homologous to those of the gamma chains than the other mouse heavy chains. Two membrane exons, separated by an 80-base intron, were identified 1.7 kb 3' to the CH4 domain of the epsilon gene and shown to conserve a hydrophobic portion similar to those of other heavy chain genes. RNA blot hybridization showed that the epsilon membrane exons are transcribed into two species of mRNA in an IgE hybridoma.

Amino Acid Sequence↗
PubMed1982

Long terminal repeat-like elements flank a human immunoglobulin epsilon pseudogene that lacks introns.

There are at least three immunoglobulin epsilon genes (C epsilon 1, C epsilon 2, and C epsilon 3) in the human genome. The nucleotide sequences of the expressed epsilon gene (C epsilon 1) and one (C epsilon 3) of the two epsilon pseudogenes were compared. The results show that the C epsilon 3 gene lacks the three intervening sequences entirely and has a 31-base A-rich sequence 16 bases 3' to the putative poly(A) addition signal, indicating that the C epsilon 3 gene is a processed gene. The C epsilon 3 gene sequence is homologous to the five separate DNA segments of the C epsilon 1 gene; namely, a segment in the 5'-flanking region (100 bases) and four exons, which are interrupted by a spacer region or intervening sequences. Long terminal repeat (LTR)-like sequences which contain TATAAA and AATAAA sequences as well as terminal inverted repeats are present in both 5'- and 3'-flanking regions. The 5' and 3' LTR-like sequences do not, however, constitute a direct repeat, unlike transposable elements of eukaryotes and retroviruses. The 3' LTR-like sequence is repetitive in the human genome, but is not homologous to the Alu family DNA. Models for the evolutionary origin of the processed gene flanked by the LTR-like sequences are discussed. The C epsilon 3 gene has a new open frame which codes potentially for an unknown protein of 292 amino acid residues.

Amino Acid Sequence↗
PubMed1982

Structure of human immunoglobulin gamma genes: implications for evolution of a gene family.

We have cloned five human immunoglobulin gamma genes from a fetal liver gene library. Four of them encode the known human immunoglobulin gamma chains gamma 1, gamma 2, gamma 3 and gamma 4. A fifth gamma gene seems to be a pseudogene. Nucleotide sequence determination demonstrates that the gamma 3 gene contains four separate hinge exons. Comparison of these hinge exons with those of the other gamma genes indicates that the first hinge exon is homologous to that of the pseudogene, and that the other three hinge exons are homologous to that of the gamma 1 gene, suggesting that the gamma 3 gene ancestor is a hybrid gene created by unequal crossing-over between the ancestral gamma 1 and psi gamma genes. Amplification of the gamma 1-type hinge exon probably followed to complete the gamma 3 gene. This hypothesis inevitably postulates the gene order 5'-gamma 1-gamma 3-psi gamma-3'. Cloning of overlapping chromosomal segments demonstrates that the gamma 2 gene is located 19 kb 5' to the gamma 4 gene. These analyses indicate that the human gamma-gene family has evolved by several types of DNA rearrangemet, including duplication of a complete gene; duplication of a hinge exon; and reassortment of exons by unequal cross-over between two adjacent genes.

Animals↗
PubMed1982

Organization of the constant-region gene family of the mouse immunoglobulin heavy chain.

We cloned overlapping DNA segments that encompass the region from the immunoglobulin JH segments to the C gamma 3 gene of BALB/c mouse. We have now cloned the entire region (about 200 kilobases) of the constant-region gene family of the immunoglobulin heavy chain, the organization of which is 5'-JH-6.5 kb-C mu-4.5 kb-C delta-55 kb-C gamma 3-34 kb-C gamma 1-21 kb-C gamma 2b-15 kb-C gamma 2a-14 kb-C epsilon-12 kb-C alpha-3'. Using these cloned DNAs, we have characterized several structural features of the constant-region gene loci. There are no other J region segments except for those at the 5' side of the C mu gene. The S region is 5' to each CH gene except for the C delta gene, and the nucleotide sequences of the S region share some homology. There is no reasonably conserved pseudogene. There are at least two species of reiterated sequences scattered in these loci. Cloning and Southern blot hybridization analyses indicate that the general organizations of the heavy-chain gene loci of BALB/c and C57BL/6 mice, which have many different serological markers, are fundamentally similar but different in the lengths of S regions. Restriction enzyme cleavage maps of the whole constant-region gene loci were constructed with respect to eight restriction endonucleases.

Animals↗
PubMed1982

Cloning of human immunoglobulin epsilon chain genes: evidence for multiple C epsilon genes.

An active human epsilon chain gene was cloned from a phage library containing partial EcoRI digests of IgE-producing myeloma DNA, using the human JH (joining) gene fragment as a probe. The epsilon chain gene clone was identified by partial nucleotide sequence determination. The germ-line constant region gene of the epsilon chain (C epsilon gene) was cloned from a human fetal liver DNA library, using the cloned epsilon chain gene as a probe. Comparative studies on the human and mouse germ-line epsilon chain genes revealed that the switch (S) sequence is more conserved than the coding sequence. Restriction endonuclease BamHI digestion of human DNA produced three C epsilon fragments of 3.0, 6.5, and 9.2 kilobases, which were named C epsilon 1, C epsilon 2, and C epsilon 3 genes, respectively. We found the three C epsilon gene fragments in all of the human DNA preparations from eleven individuals. The C epsilon gene expressed in the myeloma was identified as the C epsilon 1 gene. Because the C epsilon 2 gene is deleted from the myeloma DNA, the order of the C epsilon genes is likely to be 5'-C epsilon 2-C epsilon 1-C epsilon 3-3', assuming that all the C epsilon genes are on chromosome 14. The germ-line C epsilon 3 gene was also cloned from the myeloma DNA. Characterization of the C epsilon 3 gene revealed that it does not have the S region, suggesting that it might be a pseudogene.

Base Sequence↗
PubMed1982

A T15-idiotype-positive T suppressor hybridoma does not use the T15 VH gene segment.

The T suppressive factor (TsF) released from a T15-idiotype-positive phosphocholine (PCho)-specific T hybridoma, F18-3-4, which was formed by fusion between BALB/c T cells and BW5147 thymoma, was immunochemically characterized. TsF inhibited the in vitro induction of both IgE and IgG1 antibody responses of 2,4-dinitrophenyl keyhole limpet hemocyanin (DNP-KLH)-primed spleen cells in the presence of PCho-KLH-DNP. TsF had the ability to bind to PCho determinants and possessed T15 idiotype determinants as well as Iad products. However, we were unable to detect either the rearrangement of the T15 VH gene or the presence of T15 VH gene transcripts in hybridomas by DNA and RNA blot hybridization analyses with the T15 VH DNA probe.

Animals↗
PubMed1982

Nucleotide sequences of gene segments encoding membrane domains of immunoglobulin gamma chains.

The nucleotide sequences of the exons encoding membrane-bound IgG1, IgG2a, and IgG2b of mouse were determined and compared with the sequence of membrane-bound IgM. The sequences indicate that membrane-bound gamma chains bear an additional 71 residues at the COOH termini, including 17-residue acidic, 26-residue hydrophobic, and 28 residue hydrophilic portions. The hydrophobic portion, which seems to be anchored in the lipid bilayer of the membrane, is highly conserved among membrane-bound mu and gamma chains. We propose the presence of a membrane protein that recognizes the conserved hydrophobic segment and anchors the membrane-bound immunoglobulin. Comparison of the nucleotide sequences revealed another example of intervening sequence-mediated domain exchange around one of the membrane exons.

Amino Acid Sequence↗
PubMed1982

Relationship between the rearrangement of immunoglobulin genes, the appearance of a B lymphocyte antigen, and immunoglobulin synthesis in murine pre-B cell lines.

Eighteen Abelson virus-transformed immature B cell lines were established and immunoglobulin biosynthesis, expression of a B lymphocyte antigen detected by a monoclonal antibody, and rearrangement of immunoglobulin genes in these cell lines were studied. Only one cell line (A1) synthesized micro-chains but no light chains, and the other cell lines synthesized no detectable immunoglobulins. None of the cell lines established had detectable membrane-associated IgM. Fifteen cell lines expressed a B lymphocyte antigen on their cell surfaces. In three cell lines, however, the majority (greater than 99%) of cells did not express this antigen. Heavy chain genes were rearranged on both chromosomes in all the cell lines, although one heavy chain gene was deleted in three cell lines. In 12 of 18 cell lines, one or both kappa-chain genes were rearranged. In six cell lines, however, both kappa-chain genes remained in embryonic form; lambda-chain genes were in embryonic form in all the cell lines. These results suggested the hierarchy of Ig gene rearrangements, beginning with mu and proceeding to kappa and then to lambda. JH rearrangement was also shown to precede the appearance of a B lymphocyte antigen. In three cell lines (A1-A3), which were considered subclones derived from a single common precursor, it was suggested that one rearranged JH gene was functional, and the other was nonfunctional, indicating that allelic exclusion already operated in pre-B cells.

Abelson murine leukemia virus↗
PubMed1981

The complete nucleotide sequence of mouse immunoglobin gamma 2a gene and evolution of heavy chain genes: further evidence for intervening sequence-mediated domain transfer.

We have determined the complete nucleotide sequence (1990 base pairs) of mouse immunoglobulin gamma 2a gene, and compared it with the sequences of other gamma subclass genes so far sequenced, i.e. gamma 1 and gamma 2b genes. Divergence of the nucleotide sequence between a compared pair of the gamma genes varies extensively among different segments of the gene. For example, comparison of the gamma 2a and gamma 2b genes has revealed a remarkable homology in a long continuous segment (about 900 bases) that covers from the 3' portion of the first intervening sequence to the third intervening sequence. However, there is no particular segment of the gamma gene that is conserved universally among the three gamma genes. These findings suggest that, during their evolution, segments of the gamma genes had been scrambled between different subclass genes through recombinations within intervening sequences, thus providing further evidence for the intervening sequence-mediated domain transfer hypothesis. We have discussed several possible phylogenic trees which can explain the difference of divergence in various segments of the gamma genes.

Animals↗
PubMed1981

Ordering of mouse immunoglobulin heavy chain genes by molecular cloning.

We have determined the order of the mouse immunoglobulin gamma 1, gamma 2 b, gamma 2 a and epsilon genes by molecular cloning of overlapping chromosomal segments. The results clearly demonstrate that the order is 5'-gamma 1-(21 kilobases)-gamma 2b-(15 kilobases)-gamma 2a-(15 kilobases)-epsilon-3'. There seem to be no J regions at the 5' side of each constant region gene so far obtained except for the mu gene and these constant region genes seem to have repetitive sequences characteristic of switch (S) regions at their 5' side.

Animals↗
PubMed1981

Repetitive sequences in class-switch recombination regions of immunoglobulin heavy chain genes.

Immunoglobulin class switch involves a unique recombination event that takes place at the region 5' to each heavy chain constant region gene during B lymphocyte differentiation. Such regions that are responsible for the class-switch recombination are defined as S regions (Kataoka et al., Proc. Natl. Acad. Sci. USA 77, 919, 1980). We have cloned a rearranged gamma 2b gene from a mouse myeloma (MPC11) and compared its structure with the germ line counterparts. The rearranged gamma 2b gene contained the 5' flanking region of the gamma 3 gene (S gamma 3 region) which are linked to the 5' flanking region of the gamma 2b gene (S gamma 2b region). We have determined nucleotide sequences surrounding the recombination site of the rearranged and germ line gamma 2b genes, which include the S gamma 2b and S gamma 3 regions. Both gamma 2b and S gamma 3 regions comprise tandem repetition of conserved units of 49 bp. Similar 49 bp repeating units are also found in the previously determined sequence of the S gamma 1 region in which class-switch recombination took place in MC101 myeloma. The nucleotide sequences of the S gamma 1, S gamma 2b and S gamma 3 repeating units share significant homology with each other. The S mu region, partial nucleotide sequence of which was previously determined, contains abundant short sequences such as AGCT, TGGG and AGCTGGGG which are shared in common by repeating sequences in S gamma regions. These results suggest that the recombination responsible for class switch from mu to gamma or from a gamma to another gamma, may be facilitated directly or indirectly by homology of repeating sequences in S regions.

Animals↗
PubMed1981

Cloning of mouse immunoglobulin epsilon gene and its location within the heavy chain gene cluster.

Mouse immunoglobulin epsilon chain gene was cloned from DNA of a hybridoma producing anti-dinitrophenyl IgE, which was constructed by fusing a spleen cell of a BALB/c mouse with a variant clone of MOPC21 myeloma (IgG1 producer). Because a given active heavy chain constant region (CH) gene is linked to a heavy chain joining segment (JH) gene at its 5' side, the expressed C epsilon gene of the hybridoma was cloned from a phage library containing partial Sau3A digests of IgE hybridoma DNA by using a J gene fragment as a probe. Among 6 X 10(5) phages screened, five positive clones were obtained and three of them were identified as C epsilon gene clones by restriction mapping, Southern blot hybridization, R-loop formation, and partial nucleotide sequence determination. The determined nucleotide sequence predicted the amino acid sequence which resembles a part of the CH3 domain of human epsilon chain. The deletion profile of the C epsilon gene in various myelomas expressing different CH genes indicates that the C epsilon gene is located between the C gamma 2a and C alpha genes. The linkage (5'-epsilon-alpha-3') was directly confirmed by molecular cloning of the overlapping chromosomal segments from newborn mouse DNA.

Animals↗
PubMed1981

Structure of a rearranged gamma 1 chain gene and its implication to immunoglobulin class-switch mechanism.

An expressed gene for gamma 1 chain of MC 101 myeloma was cloned from a phage library containing partial EcoRI digests of MC 101 DNA. The cloned DNA was analyzed by restriction enzyme cleavage, Southern blot hybridization, R-loop formation, and nucleotide sequence determination. The results indicate that the expressed gamma 1 chain gene comprises at least four germline DNA segments, namely a variable-region gene, a segment of the 5' flanking region of the mu chain gene (containing J regions), a segment of the 5' flanking region of the alpha chain gene, and the gamma 1 chain gene with its flanking regions. The presence of the alpha chain gene-flanking switch (S) region (S alpha region) at the 5' side of the gamma 1 chain gene-flanking region (S gamma 1 region) indicates that the heavy chain class switch may not be mediated by stepwise linear deletion along the order of the heavy chain constant-region genes (5'-mu-gamma 3-gamma 1-gamma 2b-gamma 2a-alpha-3'). We propose a siter-chromatid exchange model that explains class switch-associated deletion of heavy chain genes by unequal crossing-over events between sister chromatids.

Base Sequence↗