FRONT COVER Strongylocentrotus purpuratus, the purple sea urchin, resting on a bed of giant kelp (Macrocystis pyrifera). Photo taken by B. Neufeld off Pt. Loma, San Diego during a collection trip to obtain gravid deep-water experimental animals for use in research in developmental biology. BACK COVER Schwann cell (red) and fibroblasts (green) identified by fluorescent antibodies in dissociated cultures of rat sciatic nerve. A REPORT FOR THE YEAR 1977-78 ON THE RESEARCH AND OTHER ACTIVITIES OF THE DIVISION OF BIOLOGY AT THE CALIFORNIA INSTITUTE OF TECHNOLOGY PASADENA, CALIFORNIA ii STAFF OF BIOLOGY 1978 Bernita K. Larsh: Compilation and typing. Elizabeth T. Hanson: Copy editing. The helpful assistance of Barbara Cathey, Jane Chacon, JoAnn Chikahiro, Geraldine Cranmer, and Lody Kempees is greatly appreciated. Pasteup: Caltech Graphic Arts. RESEARCH REPORTS Much of the research work summarized here has not yet been reported in print, in many instances because it is not yet complete. For that reason this report is not intended as a publication and should not be cited as such. Individual projects should be referred to only if specific permission to do so is obtained from the investigator responsible for the material. References are made here to published papers bearing on the projects reported. Publications by members of the Division, covering the period July 1977-June 1978, are listed separately, at the end of the research reports of each group. iii TABLE OF CONTENTS Page Introduction. • • • . • • • • . 1 Staff of Instruction and Research 9 Research Reports MOLECULAR BIOLOGY AND BIOCHEMISTRY Giuseppe Attardi - Summary • • • • • • • • • • • • • • • • • • • • • 17 1. Characterization of HeLa cell mitochondrial 7S DNA • • • • • • • • •• 17 2. Sequence analysis of the origin of replication of human mitochondrial DNA • 17 3. Transcription map of the mitochondrial DNA from HeLa cells ••••••• 19 4. Localization of promoter sites on HeLa cell mitochondrial DNA • • • • . • 19 5. Characterization of polypeptides synthesized on mitochondrial ribosomes. • 19 6. Isolation of myeloma lines secreting antibodies to human cytochrome oxidase • 20 7. Mutants of human cells deficient in mitochondrial protein synthesis • • 20 8. Mitochondrial DNA in somatic cell hybrids • • • • • • • • • • . • 20 9. Regions of homology of mitochondrial DNA from human and mouse cells 21 James Bonner - Summary. • • • • • • • • • • • • • 22 10. The role of histone acetylation in gene expression • • • 24 11. Molecular cloning of rat chromosomal DNA fragments . 25 12. Isolation of cloned ribosomal RNA coding sequences • • • • • 25 13. Analysis of rat long repetitive DNA sequences by recombinant DNA methods • 25 14. Immunochemical isolation and purification of rat albumin and et-fetoprotein mRNAs 26 15. Synthesis of full length cDNAs from purified rat albumin and a-fetoprotein mRNAs 26 16. Determination of the number of albumin and a-fetoprotein genes in rat DNA and hepatoma DNA • • • • • • • • • . • • • • . • • • • . 27 17. The nuclear RNA precursors of albumin mRNA and a-fetoprotein mRNA 27 18. Purification and mapping of slime mold ribosomal DNA •••••• 27 19. An R-loop map of rat rRNA genes isolated from total genomal DNA • 28 20. Foldback DNA in the rat genome . • • . • • • • • . • • ••• 28 21. Chromatin reconstitution • • • . • • • • . • • • • • . • • 29 22. Nucleosome structure of newly replicated chromatin •••..••• 29 23. The effect of single base mismatch on synthetic DNA-0Xl 7 4 DNA duplexes. 29 24. A comparison of mammalian and slime mold histones • • • • . • • • 30 25. Computerized analysis of thermal denaturation of DNA and chromatin • 30 Eric H. Da~n - Summary • • • • • • • • • • • • . • • • • 32 26. Studies of nucleic acid reassociation kinetics . • • • • . • • 32 27. Sequence content of the bi thorax region of the Drosophila genome. 32 28. Complexity of Drosophila egg RNA • • • • • • • . • • • • 33 29. RNA synthesis in sea urchin oogenesis • • . • • • • • . • • • • • •. 33 30. Evidence for maternally inherited poly(A) containing RNA in mouse eggs • 33 31. Cloned sea urchin DNA complementary to oocyte RNA. I. CS0088 • . 34 32. Cloned sea urchin DNA complementary to oocyte RNA. 1L PSC34 • . • 34 33. Repetitive sequence transcripts in the sea urchin oocyte . • • . •••• 34 34. Maternal mRNA localization and utilization in sea urchin development. • 35 35. Sea urchin maternal mRNA sequences also synthesized during embryogenesis 35 36. Nonmaternal messenger RNA sequences in sea urchin embryos • • • . • • 36 37. Sea urchin embryo mRNA sequences in the nuclear RNA of adult tissues .• 36 38. Repetitive sequence transcripts in sea urchin nuclear RN As. • . • • • 36 39. Repeated sequences in the vicinity of expressed single copy sea urchin DNA 37 40. Primary structure of repetitive DNA sequences in the sea urchin genome . 37 41. Selection of cloned sea urchin DNA coding for mRNA .••• 37 42. Single copy DNA sequence polymorphism. • • • • • . • 38 43. Evolutionary change in sea urchin repetitive DNA • • . • . 38 44. Sequence organization of individual repeated sequence families • 39 45. Single copy DNA sequence differences among related sea urchins 39 William J. Dreyer - Summary • • • • • • • • • • • • • • • • • • 40 46. Fractionation and analysis of proteins displayed on the surface of tumor cells . 41 4 7. Identification of cell-surface molecules that react with anti-tumor serum 41 48. Studies of the structure and function of a family of cell-surface recognition molecules . 41 49. Establishment of an advanced laboratory for studies of proteins • • • • • •• 42 50. A new, extraordinarily sensitive analytical system for research and medicine • 42 51. Is developmentally programmed gene splicing used only in the immune system? 43 iv Leroy E. Hood - Summary • • • • . • . • • • . 44 52. Sequence analysis of a closely-related set of mouse VK regions 45 53. Sequence studies of a-1,3 dextran-binding immunoglobulins .. 46 54. Sequence studies of levan-binding immunoglobulins • • . • 46 55. Construction of immunoglobulin cDNA plasmids .•..••• 47 56. The construction of mouse DNA libraries and the characterization of immunoglobulin genes. 47 57. Determination of the structure of mouse lgM • • • • . • • • • . • 47 58. Comparison of the structure of soluble and membrane-bound mouse lgM 48 59. Characterization of a putative T cell receptor . 48 6 0. Analysis of the diversity of mouse transplantation antigens • 49 61. Site of polymorphism in mouse H-2 transplantation antigens 49 62. Direct sequence studies of mouse transplantation antigens • 49 63. Biochemical analysis of mutant H-2 antigens • . • • 49 64. Ia on cells in the epidermis • • . • • . 50 65. Chemical characterization of Ia antigens of the mouse major histocompatibility complex (MHC). • . • • . • • . • • • • . • 50 66. Chemical characterization of la antigens from murine tumor cells • 51 67. Development of automated protein microsequencing techniques . 51 Norman H. Horowitz - Summary. 52 68. Laboratory simulations of Viking results . • • • . • • . 52 69. Siderophore binding to Neurospora cytoplasmic membranes . 53 70. Experiments with a siderophore-deficient mutant of Neurospora. 53 71. Siderophores of Aspergillus and Penicillium. 54 Tom Maniatis - Summary. • • • . • • 54 72. Construction of libraries of eukaryotic DNA 55 7 3. Characterization of linked 13-globin genes in rabbit 56 7 4. Restriction mapping and nucleotide sequence analysis of cloned 13-globin genes 56 75. Other rabbit globin genes ..•••...•••• 56 76. Isolation and characterization of human globin genes •..• 57 77. Y chromosome genes of Drosophila ..••...•••. 57 7 8. The alcohol dehydrogenase gene of Drosophila melanogaster 57 Herschel K. Mitchell - Summary. • • • . • 58 79. Heat shock phenocopies in Drosophila • • . • 58 8 0. Heat shock and protein synthesis in Drosophila 59 81. Purification of a heat shock protein . • • 59 82. Mapping of the coding region for two heat shock proteins. 60 83. Development and protein synthesis of the flight muscle of Drosophila melanogaster 60 84. The aromatic cross-link in the cuticle of Drosophila 60 85. Teratogens and phenocopies in Drosophila • . • • • • . • • • . • • • • 61 James H. Strauss Jr. - Summary. 61 86. Amino terminal sequence analysis of the structural proteins of Sindbis virus. 62 87. Amino acid compositions of the structural proteins of Sindbis virus 62 88. Production and analysis of cyanogen bromide fragments of Sindbis proteins 63 89. Host polypeptides present in purified HR and ts103 8indbis virions • • • 63 90. Growth and release of several alpha-viruses in chick and BHK cells • • 63 91. Characterization of E3 in the 8indbis virus infection • • . • • • • • • 64 92. Investigation of the cleavages of the structural proteins of 8indbis virus 64 93. Structural studies on 8indbis virus . • . • • • • . • • • • . • • 64 94. Antibodies against Sindbis structural proteins • • • • • . • • • • • , 65 95. Biophysical studies on the cyclization of 498 RNA of 8indbis virus ••• 65 96. Location of 5-methylcytidine residues in 8indbis virus RNAs • • • • . 66 97. The determination of the 5'-terminal and 3'-terminal sequences of 8indbis 498 and 268 RNAs 66 CELLULAR BIOLOGY AND BIOPHYSICS Charles J. Brokaw - Summary. • • . • • • . • • . • . • • • • . • • • • • • • • • • 69 98. Flagellar hydrodynamics: A comparison between resistive force theory and slender-body theory • 69 99. Inhibition of flagellar movement by components of the ATP system 69 100. Reactivation of demembranated Ciona spermatozoa • • . • • 69 101. Flagellar