1994

M e t h o d s in Y e a s t G e n e t i c s ............. A Cold Spring Harbor Laboratory Course Manual, 1994 Edition By Chris Kaiser, Massachusetts Institute of Technol- ogy; Susan Michaelis, Johns Hopkins Medical School; Aaron Mitchell, Columbia University Col- lege of Physicians & Surgeons

The Cold Spring Harbor Yeast Genetics Course, founded in 1970, evolves continuously to present a combination of standard methods and the latest tech- niques in yeast biology. This is the latest edition of the course manual, which incorporates a variety of new techniques with revised experiments and the es- sential elements from the 1987 and 1990 publica- tions. The book remains the primary source of techni- cal guidance for anyone using yeast, either as a new- comer or as an established investigator.

CONTENTS Introduction Genetic Nomenclature Experiments Looking at Yeast Cells; I. Isolation and Characterization of Auxotrophic, Temperature-sensitive, and UV-sensitive Mutants; II. Meiotic Mapping; III. Mitotic Recombination: Loss of Heterozygosity and Mitotic Mapping; IV. Trans- formation of Yeast; V. Complementation and Random Spore Analysis of Histidine Auxotrophs; VI. Gene Re- placement; VII. Isolation of ras2 Suppressors; VIII. The Formation of Zygotes: Mating and Karyogamy; IX. Clon- ing of Yeast Genes by Complementation; X. lacZ Gone Fu- sion Expression in Yeast; XI. lmmunofluorescent Staining

of Yeast Cells; XII. Eiectrophoretic Karyotyping and Gene Mapping Using Pulsed-field Gel Electrophoresis Techniques and Protocols 1. Yeast Transformation; 2. Lithium Acetate Yeast Trans- formation; 3. Yeast Colony Hybridization; 4. Yeast DNA Isolations: a. Yeast DNA Miniprep (40 ml), b. Yeast DNA Miniprep (5 ml), c. A Ten-minute DNA Prep from Yeast, d. Preparation of Yeast Genomic DNA (CsCi Method); 5. Yeast Protein Extracts; 6. Yeast RNA Isolation; 7. Northern Analysis: Formaldehyde Agarose Gel, Blotting, and Hybridization to Filters; 8. Alkaline Southern Blotting Pro- cedure; 9. Scoring Killer Factor; 10. Hydroxylamine Mutagenesis of Plasmid DNA; 11. Assay of I]-Galacto- sidase in Yeast; 12. Transformation of Bacteria (CaCI 2 Method); 13. E. coli Plasmid DNA Miniprep; 14. Prepara- tion of Chromosome-sized Yeast DNA Molecules in Solid Agarose; 15. Mapping Cloned DNA Segments by Chromo- some "Fragmentation"; 16. Running Gels Using the CHEF Apparatus; 17. Running Gels Using the OFAGE Apparatus; 18. Yeast Vital Stains; 19. Yeast lmmunofluorescence with Antibodies; 20. Actin Staining in Fixed Cells; 21. Modified Lithium Acetate Yeast Transformation; 22. PCR Analysis of Genotype Appendices A. Media; B. Stock Preservation; C. Yeast Genetic Map; D. Grids; E. Electrophoretic Karyotypes of Strains for Southern Blot Mapping; F. Strains

1994, 202 pp., illus., appendices ISBN 0-87969-451-3 Plastic comb binding $49

Reader Service No. 277

GENES

DEVELOPMENT

VOLUME 8 NUMBER 24 PAGES 2939-3110 DECEMBER 15, 1994

E D I T O R I A L B O A R D

I. Adams {Melbourne, Australia) I. Beckwith {Boston, USA) T. Cech {Boulder, USA) P. Chambon {Strasbourg, France) N.-H. Chua {New York, USAI E. Coen {Norwich, UKI J. Coffin {Boston, USA) S. Courmeidge {Heidelberg, FRG) E. De Robertis (Los Angeles, USA) R. Evans (La Jolla, USA) G. Fink (Cambridge, USA) P. Goodfellow {Cambridge, UK) S. Gottesman (Bethesda, USA) T. Graf {Heidelberg, FRG) C. Gross (San Francisco, USA) R. Grosschedl (San Francisco, USA) F. Grosveld {Rotterdam, The Netherlands) M. Groudine {Seattle, USA) L. Guarente {Cambridge, USAI R. Harland IBerkeley, USA) E. Harlow {Charlestown, USA) W. Herr {Cold Spring Harbor, USA) J. Hodgkin {Cambridge, UK) R. Horvitz {Cambridge, USA) P. Ingham {London, UK) T. Jessell {New York, USAJ

N. Jones {London, UK} J. Kadonaga (La lolla, USA) R. Lehmann (Cambridge, USA) M. Levine (San Diego, USA) D. Livingston {Boston, USA) R. Losick {Cambridge, USA) J. Manley {New York, USA) W. McGinnis {New Haven, USA) S. McKnight (South San Francisco, USA) A. McMahon {Cambridge, USA) P. Nurse (London, UK) C. Nfisslein-Volhard (Tubingen, FRG) R. Palmiter {Seattle, USA) L. Parada (Dallas, USA) C. Prives (New York, USA) G. Rubin (Berkeley, USA) U. Schibler {Geneva, Switzerland) D. Solter (Freiburg, FRG) P. Soriano {Seattle, USAI J. Steitz (New Haven, USA) T. Taniguchi {Osaka, Japan) S. Tilghman (Princeton, USA) R. Tjian (Berkeley, USA) E. Wagner (Vienna, Austria) V. Walbot {Stanford, USA) M. Wigler (Cold Spring Harbor, USA)

Editors T. Grodzicker {Cold Spring Harbor) N. Hastie {Edinburgh)

Managing Editor J. Cuddihy (Cold Spring Harbor)

Editorial~Production N. Dumser, Technical Editor V. Nicolette, Production Editor L. Olsewski, Editorial Secretary

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GENES & DEVELOPMENT (ISSN 0890-9369) is published semimonthly for $450 (institutional), $115 (individual making personal paymentl, $159 (Genet- ical Society of Great Britain members) by Cold Spring Harbor Laboratory Press, Bungtown Road, Cold Spring Harbor, New York 11724, in associ- ation with the Genetical Society of Great Britain. Second-class postage is paid at Cold Spring Harbor and additional mailing offices. POSTMAS- TER: Send address changes to Cold Spring Harbor Laboratory Press, POB 100, 1 Bungtown Rd., Cold Spring Harbor, New York 11724-2203. Subscription Price Orders may be sent to Cold Spring Harbor Labora- tory Press, Fulfillment Department, 10 Skyline Drive, Plainview, NY 11803-9729. Telephone: Continental US except NY State, 1-800-843- 4388; all other locations, 516-349-1930/1931/1932. FAX 516-349-1946. Volume 8, 1994, $450, U.S. institutional; $540, R.O.W. institutional. Personal subscription rate: $115, U.S.; $205, R.O.W. Price includes sur- face postage for U.S. and airlift for R.O.W. Genetical Society members, $159.00. All subscriptions are entered for the calendar year and must be prepaid. Personal subscriptions must be prepaid by personal check, credit card, or money order. All checks must be for US dollars and drawn on a US bank. Genetical Society members may also subscribe by check, payable to the Genetical Society, for £115 (includes airlift}. Send to: Dr.

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Claims for missing issues must be received within 4 months of issue date. Change of address Please enclose recent mailing label with address change; allow 4 weeks. Advertising To advertise in Genes & Development, contact Nancy Kuhle, Advertising Manager, Cold Spring Harbor Laboratory Press, Cold Spnng Harbor, New York 11724-2203; telephone 516-367-8351. Photo Copy Authorization to photocopy items for internal or personal use, or the internal or personal use of specific clients, is granted by Cold Spring Harbor Laboratory Press for libraries and other users registered with the Copyright Clearance Center (CCC] Transactional Reporting Service, provided that the base fee of $5.00 per copy is paid directly to CCC, 21 Congress St., Salem, MA 01970 (0890-9369/94 $5.00 + 0.). This consent does not extend to other kinds of copying, such as copying for general distribution for advertising or promotional purposes, for creating new collective works, or for resale. Copyright © 1994 by Cold Spring Harbor Laboratory Press

Contents GENES & DEVELOPMENT December 15, 1994

Research papers Growth suppression by p18, a p16 nvK¢/MTsl- and pl4rNg4WMTSZ-related CDK6 inhibitor, correlates with wild-type pRb function Kun-Liang Guan, Christopher W. Jenkins, Yan Li, Michael A. Nichols, Xiaoyu Wu, Christine L. O'Keefe, A. Gregory Matera, and Yue Xiong

2939

Loss of the imprinted IGF2/cation-independent mannose 6-phosphate receptor results in fetal overgrowth and perinatal lethality Michele M.H. Lau, Claire E.H. Stewart, Ziying Liu, Harshida Bhatt, Peter Rotwein, and Colin L. Stewart

2953

Colorectal hyperplasia and inflammation in keratin 8-deficient FVB/N mice H41~ne Baribault, Jocelyn Penner, Renato V. Iozzo, and Marcia Wilson-Heiner

2964

Elements of a single MAP kinase cascade in Saccharomyces cerevisiae mediate two developmental programs in the same cell type: mating and invasive growth Radclyffe L. Roberts and Gerald g. Fink

2974

RNA localization along the anteroposterior axis of the Drosophila oocyte requires PKA-mediated signal transduction to direct normal microtubule organization Mary Ellen Lane and Daniel Kalderon

2986

JNK2 contains a specificity-determining region responsible for efficient c-Jun binding and phosphorylation Tuula Kallunki, Bing Su, Igor Tsigelny, Hayla K. Sluss, Benoit D~rijard, George Moore, Roger Davis, and Michael Karin

2996

Dynamic association of proteins with the pre-mRNA branch region Andrew M. MacMillan, Charles C. Query, Charles R. Allerson, Swaine Chen, Gregory L. Verdine, and Phillip A. Sharp

3008

Ribonuclease E provides substrates for ribonuclease P-dependent processing of a polycistronic mRNA Pietro Alifano, Flavia Rivellini, Claudia Piscitelli, Cecilia M. Arraiano, Carmelo B. Bruni, and M. Stella Carlomagno

3021

fgfr-1 is required for embryonic growth and mesodermal patterning during mouse gastrulation Terry P. Yamaguchi, Kendraprasad Harpal, Mark Henkemeyer, and Janet Rossant

3032

Murine FGFR-1 is required for early postimplantation growth and axial organization Chu-Xia Deng, Anthony Wynshaw-Boris, Michael M. Shen, Cathie Daugherty, David M. Ornitz, and Philip Leder

3045

(continued)

Location, structure, and function of the target of a transcriptional activator protein Hong Tang, Konstantine Severinov, Alex Goldfarb, David Fenyo, Brian Chait, and Richard H. Ebright

3058

Retinoid-dependent in vitro transcription mediated by the RXR/RAR heterodimer Rafael Valc.4rcel, Herbert Holz, Custodia Garcia Jim~nez, Domingo Barettino, and Hendrik G. Stunnenberg

3068

Reviewers, Volume 8 3080

Advertisers, Volume 8 3083

Author Index, Volume 8 3084

Subject Index, Volume 8 3088

Covet Drosophila PKA and A/P axis determination. (For details, see Lane and Kalderon, p. 2986.)