Analysis of the effect of a strain-specific alteration in the upstream octamer region on transcription of a mouse V kappa Ser light chain gene.
Previous studies had shown that several phenotypic markers expressed by several strains of mice (C58, AKR, PL, RF) involve a light chain group, called V kappa Ser, encoded by a single germ-line gene (Igk-VSera). Most other inbred strains (e.g., BALB/c) contain an allele (Igk-VSerb) which differs from Igk-VSera both in coding regions and in an upstream octamer region known to be important for transcription. Since no evidence for expression of the Igk-VSerb gene product has been observed, experiments were undertaken to determine whether the alteration in the regulatory octamer region of BALB/c might have rendered it defective for transcription. The upstream octamer-containing region of a cloned functional V kappa Ser gene expressed by the C.C58 myeloma M75 was replaced by the corresponding region from BALB/c or deleted entirely. Constructs were transfected into J558L cells and quantity of transcription and site of transcription initiation were compared. Results suggest that the BALB/c octamer (CTTTGCGT), which differs at two of eight nucleotides from the consensus octamer sequence (ATTTGCAT), is fully functional in transcription initiation. This is consistent with results of S1 nuclease protection experiments which indicate the presence of small amounts of correctly initiated V kappa Ser-related RNA in BALB/c spleen.
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D005803:Genes, Immunoglobulin; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D007145:Immunoglobulin kappa-Chains / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D012313:RNA / Q000032:analysis; D012045:Regulatory Sequences, Nucleic Acid; D013045:Species Specificity; D013154:Spleen; D014158:Transcription, Genetic; D014162:Transfection
D007135:Immunoglobulin Variable Region; D007145:Immunoglobulin kappa-Chains; D012313:RNA