Abstract
The primary sequence and higher order structures of a G+C-rich satellite DNA of the Bermuda land crab Gecarcinus lateralis have been described previously. The repeat unit of the satellite is approximately 2.1 kb. In exploring a possible function for this satellite, we asked whether it is transcribed. As a probe for transcripts, we used a segment of DNA amplified from a 368 bp EcoRI fragment from the very highly conserved 3′ end of the satellite DNA. During polymerase chain reaction (PCR) amplification, the probe was simultaneously either radiolabeled or biotinylated. Tissue- and stage-specific transcripts were observed when blots of poly(A)+ mRNAs recovered from polysomes isolated from crab tissues [including midgut gland (hepatopancreas), limb bud, and claw muscle] were probed with the satellite DNA fragment. The presence of satellite transcripts in polysomal mRNAs is strong evidence that the transcripts had reached the cytoplasm. To corroborate the presence of transcripts in the cytoplasm, we investigated in situ hybridization of satellite probes with RNAs in tissue sections. Biotinylated satellite DNA probes were applied to sections of midgut gland, limb bud papilla, ovary, or testis of anecdysial crabs. Retention of RNAs in tissue sections was improved by UV-irradiation prior to hybridization. Transcripts were abundant in the cytoplasm of all tissues except testis. Sections of crab midgut gland treated with RNase A prior to hybridization and sections of mouse pancreatic tumor served as controls; neither showed any signals with the probe.
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Baldwin L, Macgregor HC (1985) Centromeric satellite DNA in the newts Triturus cristatus karelinii and related species: its distribution and transcription on lampbrush chromosomes. Chromosoma 92:100–107
Barsacchi-Pilone G, Batistoni R, Andronico F, Vitelli L, Nardi I (1986) Heterochromatic DNA in Triturus (Amphibia, Urodela). I. A satellite DNA component of pericentric C-bands. Chromosoma 93:435–446
Baumlein H, Wobus U, Gerbi SA, Kafatos FC (1982a) The basic repeat unit of a Chironomus Balbiani ring gene. Nucleic Acids Res 13:3893–3904
Baumlein H, Wobus U, Gerbi SA, Kafatos FC (1982b) Characterization of a 249-bp tandemly repetitive, satellite-like repeat in the translated portion of Balbiani ring c of Chironomus thummi. EMBO J 1:641–647
Biesiot PM, Wang SY, Skinner DM (1987) Discrete segments of a complex eukaryotic satellite are dispersed in a crab genome. J Cell Biol 105:1365
Bonaccorsi S, Pisano C, Puoti F, Gatti M (1988) Y chromosome loops in Drosphila melanogaster. Genetics 120:1015–1034
Bonaccorsi S, Gatti M, Pisano C, Lohe A (1990) Transcription of a satellite DNA on two Y chromosome loops of Drosophila melanogaster. Chromosoma 99:260–266
Bonnewell V, Fowler RF, Skinner DM (1983) An inverted repeat borders a fivefold amplification in satellite DNA. Science 221:862–865
Bromley S, Gall JG (1987) Transcription of the histone loci on lampbrush chromosomes of the newt Notophthalmus viridescens. Chromosoma 95:396–402
Brutlag DL (1980) Molecular arrangement and evolution of heterochromatic DNA. Annu Rev Genet 14:121–144
Church GM, Gilbert W (1984) Genome sequencing. Proc Natl Acad Sci USA 81:1991–1995
Clemens MJ (1984) Purification of eukaryotic messenger RNA. In: Hames BD, Higgins SJ (eds) Transcription and translation: a practical approach. IRL Press, Oxford, England, pp 211–230
Degelmann A, Hollenberg CP (1981) A structural analysis of Balbiani ring DNA sequences in Chironomus tentans. Chromosoma 83:295–313
Demas S, Duronslet M, Wachtel S, Caillouet C, Nakamura D (1990) Sex-specific DNA in reptiles with temperature sex determination. J Exp Zool 253:319–324
Diaz MO, Barsacchi-Pilone G, Mahon KA, Gall JG (1981) Transcripts from both strands of a statellite DNA occur on lampbrush chromosome loops of the newt Notophthalmus. Cell 24:649–659
Epstein LM, Mahon KA, Gall JG (1986) Transcription of a satellte DNA in the newt. J Cell Biol 103:1137–1144
Fowler RF, Bonnewell V, Spann MS, Skinner DM (1985) Sequences of three closely related variants of a complex satellite DNA diverge at specific domains. J Biol Chem 260:8964–8972
Fowler RF, Stringfellow LA, Skinner DM (1988) A domain that assumes a Z-conformation includes a specific deletion in some cloned variants of a complex satellite. Gene 71:165–176
Gallyas F, Merchenthaler I (1988) Copper-H2O2 oxidation strikingly improves silver intensification of the nickel-diaminobenzidine (Ni-DAB) end-product of the peroxidase reaction. J Histochem Cytochem 36:807–810
Graham DE, Skinner DM (1973) Homologies of repetitive DNA sequences among Crustacea. Chromosoma 40:135–152
Hackstein JHP, Leoncini O, Beck H, Peelen G, Hennig W (1982) Genetic fine structure of the Y chromosome of Drosophila hydei. Genetics 101:257–277
Hennig W (1985) Y chromosome function and spermatogenesis in Drosophila hydei. J Mol Biol 38:1–13
Hennig W (1993) Conventional protein coding genes in the Drosophila Y chromosome: is the puzzle of the fertility gene function solved? Proc Natl Acad Sci USA 90:10904–10906
Hochstenbach R, Wilbrink M, Suijkerbuijk R, Hennig W (1993a) Localization of the lampbrush loop pair 3 ‘Nooses’ on the Y chromosome of Drosophila hydei by fluorescence in situ hybridization. Chromosoma 102:546–552
Hochstenbach R, Potgens A, Meijer H, Dijkhof R, Knops M, Schouren K, Hennig W (1993b) Partial reconstruction of the lampbrush loop 3 ‘Nooses’ on the Y chromosome of Drosophila hydei. Chromosoma 102:526–545
Huijser P, Hennig W (1987) Ribosomal DNA-related sequences in a Y chromosomal lampbrush loop of Drosophila hydei. Mol Gen Genet 206:441–451
Huijser P, Hennig W, Dijkhof R (1987) Poly(dC-dA/dG-dT) repeats in the Drosophila genome: a key function for dosage compensation and position effects. Chromosoma 95:209–215
Huijser P, Kirchhoff C, Lankenau D-H, Hennig W (1988) Retrotransposon-like sequences are expressed in Y chromosomal lampbrush loops of Drosophila hydei. J Mol Biol 203:689–697
Huijser P, Beckers L, Top B, Hermans M, Sinke R, Hennig W (1990) Poly[d(C-A)]poly[d(G-T)] is highly transcribed in the testes of Drosophila hydei. Chromosoma 100:48–55
Jamrich M, Warrior R, Steele R, Gall JG (1983) Transcription of repetitive sequences on Xenopus lampbrush chromosomes. Proc Natl Acad Sci USA 80:3364–3367
Kirchhoff C (1988) GATA tandem repeats detect minisatellite regions in blowfly DNA (Diptera: Calliphoridae). Chromosoma 96:107–111
LaMarca ME, Allison DP, Skinner DM (1981) Irreversible denaturation mapping of a pyrimidine-rich domain of a complex satellite DNA. J Biol Chem 256:6475–6479
Lifschytz E, Hareven D, Azriel A, Brodsly H (1983) DNA clones and RNA transcripts of four lampbrush loops from the Y chromosome of Drosophila hydei. Cell 32:191–199
Moyd MA, Fields MJ, Thorgaard GH (1989) Bkm minisatellite sequences are not sex associated but reveal DNA fingerprint polymorphisms in rainbow trout. Genome 32:865–868
Macgregor HC, Sessions SK (1986) The biological significance of variation in satellite DNA and heterochromatin in newts of the genus Triturus: an evolutionary perspective. Philos Trans R Soc Lond [Biol] 312:243–259
Miklos GLG, Mattnaei KL, Reed KC (1989) Occurrence of the (GATA)n sequences in vertebrate and invertebrate genomes. Chromosoma 98:194–200
Mykles DL, Skinner DM (1982) Molt cycle-associated changes in calcium-dependent proteinase activity that degrades actin and myosin in crustacean muscle. Dev Biol 92:386–397
Pisano C, Bonaccorsi S, Gatti M (1993) The kl-3 loop of the Y chromosome of Drosophila melanogaster binds a tektin-like protein. Genetics 133:569–579
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Singh L, Jones KW (1986) Bkm sequences are polymorphic in humans and are clustered in pericentric regions of various acrocentric chromosomes including the Y. Hum Genet 73:304–308
Singh L, Phillips C, Jones KW (1984) The conserved nucleotide sequences of Bkm, which define Sxr in the mouse, are transcribed. cell 36:111–120
Skinner DM (1962) The structure and metabolism of a crustacean integumentary tissue during a molt cycle. Biol Bull 123:635–647
Skinner DM (1966) Breakdown and reformation of somatic muscle during the molt cycle of the land crab, Gecarcinus lateralis. J Exp Zool 163:115–124
Skinner DM (1967) Satellite DNAs in the crabs Gecarcinus lateralis and Cancer pagurus. Proc Natl Acad Sci USA 58:103–110
Skinner DM (1977) Satellite DNA's. BioSciences 27:790–796
Skinner DM, Graham DE (1970) Molting in land crabs: stimulation by leg removal Science 169:383–385
Spohr G, Reith W, Sures I (1981) Organization and sequence analysis of a cluster of repetitive DNA elements from Xenopus laevis. J Mol Biol 151:573–592
Stringfellow LA, Fowler RF, LaMarca ME, Skinner DM (1985) Demonstration of remarkable sequence divergence in variants of a complex satellite DNA by molecular cloning. Gene 38:145–152
Tiersch TR, Simco BA, Davis KB, Wachtel SS (1992) Molecular genetics of sex determination in channel catfish: studies on SRY, ZFY, Bkm, and human telomeric repeats. Biol Reprod 47:185–192
Trapitz P, Wlaschek M, Bunemann H (1988) Structure and function of Y chromosomal DNA: II. Analysis of lampbrush loop associated transcripts in nuclei of primary spermatocytes of Drosophila hydei by in situ hybridization using asymmetric RNA probes of four different families of repetitive DNA. Chromosoma 96:159–170
Trapitz P, Glatzer KH, Bunemann H (1992) Towards a physical map of the fertility genes on the heterochromatic Y chromosome of D. hydei: Families of repetitive sequences transcribed on the lampbrush loops 3 ‘Nooses’ and 3 ‘Threads’ are organized in extended clusters of several hundred kilobases. Mol Gen Genet 235:221–234
Varadaraj K, Skinner DM (1994) Denaturants or cosolvents improve the specificity of PCR amplification of a G+C-rich DNA using genetically engineered DNA polymerase. Gene 140:1–5
Varadaraj K, Wang SY, Skinner DM (1990) Clones in a crab regenerating-limb cDNA library contain segments of the repeat unit of a complex satellite. J Cell Biol 111:2847
Varadaraj K, Kumari SS, Skinner DM (1993) α-Tubulin genes in a land crab, Gecarcinus lateralis. Mol Biol Cell 4:1518
Varley JM, Macgregor HC, Nardi I, Andrews C, Erba HP (1980) Cytological evidence of transcription of highly repeated DNA sequences during the lambrush stage in Triturus cristatus carnifex. Chromosoma 80:289–307
Varley JM, Macgregor HC, Barnett L (1990) Characterisation of a short, highly repeated and centromerically localised DNA sequence in crested and marbled newts of the genus Triturus. Chromosoma 100:15–31
Vogt P, Hennig W (1986a) Molecular structure of the lampbrush loops nooses of the Y chromosome of Drosophila hydei: I. The Y chromosome-specific repetitive DNA sequence family ay1 is dispersed in the loop DNA. Chromosoma 94:449–458
Vogt P, Hennig W (1986b) Molecular structure of the lampbrush loops nooses of the Y chromosome of Drosophila hydei: II. DNA sequences with homologies to multiple genomic locations are major constituents of the loop. Chromosoma 94:459–467
Vogt P, Hennig W, Siegmund I (1982) Identification of cloned Y chromosomal DNA sequences from a lampbrush loop of Drosophila hydei. Proc Natl Acad Sci USA 79:5132–5136
Walker PMB (1971) Origin of satellite DNA. Nature 229:306–308
Wang SY, Biesiot PM, Skinner DM (1988) Transcription of a complex satellite DNA. J Cell Biol 107:4252
Wu Z, Murphy C, Gall JG (1986) A transcribed satellite DNA from the bullfrog Rana catesbeiana. Chromosoma 93:291–297
Zeller R (1989) In situ hybridization and immunohistochemistry. In: Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (eds) Current protocols in molecular biology, vol 2. Greene Publishing Associates and Wiley-Interscience, New York, pp 14.1.1–14.1.8
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Varadaraj, K., Skinner, D.M. Cytoplasmic localization of transcripts of a complex G+C-rich crab satellite DNA. Chromosoma 103, 423–431 (1994). https://doi.org/10.1007/BF00362287
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DOI: https://doi.org/10.1007/BF00362287